Methods for quantifying inter-alpha inhibitor proteins

Abstract

Described herein are methods for quantifying lAIP levels in a sample (e.g., from a subject) using lAIP ligand-based assays. Also disclosed are methods for measuring lAIP-IAIP ligand complexes, and methods of evaluating, monitoring, and treating subjects using the aforementioned lAIP quantification methods.

1 . A method for quantifying inter-alpha inhibitor protein (IAIP) in a sample from a subject comprising:
 a) contacting the sample with a binding agent to produce an IAIP-binding agent complex, wherein the binding agent is bound to a support;   b) contacting the IAIP-binding agent complex with a detection agent; and   c) detecting an amount of the detection agent bound to the IAIP-binding agent complex to quantify IAIP in the sample.   
     
     
         2 . The method of  claim 1 , wherein the IAIP is intact IAIP. 
     
     
         3 . The method of  claim 1  or  2 , wherein the binding agent is an IAIP ligand that binds to IAIP. 
     
     
         4 . The method of  claim 1  or  2 , wherein the binding agent is an antibody that specifically binds to IAIP. 
     
     
         5 . The method of any one of  claims 1  to  4 , wherein the detection agent comprises an IAIP ligand. 
     
     
         6 . The method of  claim 5 , wherein the detection agent further comprises an antibody that binds to the IAIP ligand detection agent. 
     
     
         7 . The method of any one of  claims 1  to  4 , wherein the detection agent is an antibody that specifically binds to IAIP. 
     
     
         8 . The method of  claim 1 , wherein the IAIP is in an IAIP-IAIP ligand complex. 
     
     
         9 . The method of  claim 8 , wherein the binding agent is an IAIP ligand that binds to IAIP. 
     
     
         10 . The method of  claim 9 , wherein the IAIP ligand of the IAIP-IAIP ligand complex is different from the binding agent. 
     
     
         11 . The method of  claim 8 , wherein the binding agent is an antibody that binds to the IAIP ligand of the IAIP-IAIP ligand complex. 
     
     
         12 . The method of  claim 8 , wherein the binding agent is an antibody that specifically binds to IAIP of the IAIP-IAIP ligand complex. 
     
     
         13 . The method of any one of  claims 8  to  12 , wherein the detection agent comprises an IAIP ligand that binds to IAIP. 
     
     
         14 . The method of  claim 13 , wherein the detection agent further comprises an antibody that binds to the IAIP ligand detection agent. 
     
     
         15 . The method of  claim 13  or  14 , wherein the IAIP ligand of the IAIP-IAIP ligand complex is different from the IAIP ligand detection agent. 
     
     
         16 . The method of any one of  claims 8  to  12 , wherein the detection agent is an antibody that binds to the IAIP ligand of the IAIP-IAIP ligand complex. 
     
     
         17 . The method of any one of  claims 8  to  12 , wherein the detection agent is an antibody that specifically binds to IAIP of the IAIP-IAIP ligand complex. 
     
     
         18 . The method of any one of  claims 4 ,  6 ,  7 ,  11 ,  12 ,  14 ,  16 , and  17 , wherein the antibody is a monoclonal antibody. 
     
     
         19 . The method of any one of  claims 4 ,  7 ,  12 , and  17 , wherein the antibody is MAb 69.26 or MAb 69.31. 
     
     
         20 . The method of any one of  claims 3 ,  5 ,  6 , and  8  to  17 , wherein the IAIP ligand is selected from the group consisting of endotoxin (LPS), heparin, a histone, hyaluronic acid, vitronectin, fibronectin, laminin, tenascin C, aggrecan, von Willebrand Factor, pentraxin-3 (PTX3), TNF-stimulated gene-6 (TSG-6), factor IX, a complement component, factor XIIIa, and tissue transglutaminase. 
     
     
         21 . The method of  claim 28 , wherein the complement component is C1q, C2, C3, C4, C5, C6, C8, properdin, or factor D. 
     
     
         22 . The method of any one of  claims 1  to  21 , wherein the detection agent comprises a label. 
     
     
         23 . The method of  claim 22 , wherein the label is biotin, an enzyme, an enzyme substrate, a radiolabel, a luminescent compound, colloidal gold, a particle, or a fluorescent dye. 
     
     
         24 . The method of any one of  claims 1  to  23 , wherein the support is a plate, a particle, a nanoparticle, a resin, a membrane, a biochip, a container, a test strip, or a bead. 
     
     
         25 . The method of any one of  claims 1  to  24 , wherein the method further includes a wash step between steps a) and b). 
     
     
         26 . The method of any one of  claims 1  to  25 , wherein the method further includes a wash step between steps b) and c). 
     
     
         27 . The method of any one of  claims 1  to  26 , wherein the method further includes a blocking step prior to step a) or step b). 
     
     
         28 . The method of any one of  claims 1  to  27 , wherein the contacting in step a) and/or b) is performed at a pH of about 7.0 to about 3.5. 
     
     
         29 . The method of  claim 28 , wherein the pH is about 5.0 to about 3.5. 
     
     
         30 . The method of  claim 29 , wherein the pH is about 4.0. 
     
     
         31 . The method of any one of  claims 1  to  30 , wherein the sample is a fluid. 
     
     
         32 . The method of  claim 31 , wherein the fluid is blood, plasma, serum, urine, cerebrospinal fluid, synovial fluid, amniotic fluid, interstitial fluid, follicular fluid, peritoneal fluid, bronchoalveolar lavage fluid, breast milk, sputum, lymph, bile, or tissue homogenate. 
     
     
         33 . The method of any one of  claims 1  to  32 , wherein the subject is a human subject. 
     
     
         34 . The method of  claim 33 , wherein the subject has been identified as having or at risk of developing an inflammatory disease or condition or an infection. 
     
     
         35 . The method of  claim 33 , wherein the subject has not been identified as having or at risk of developing an inflammatory disease or condition or an infection. 
     
     
         36 . The method of  claim 33 , wherein the method is performed before, after, or concurrent with diagnosis of the subject as having or at risk of an inflammatory disease or condition or an infection. 
     
     
         37 . The method of  claim 33  or  34 , wherein the method is performed substantially concurrent with treatment of the subject for an inflammatory disease or condition or an infection. 
     
     
         38 . The method of any one of  claims 33  to  36 , wherein the method is performed prior to treatment of the subject for an inflammatory disease or condition or an infection. 
     
     
         39 . The method of  claim 33  or  34 , wherein the method is performed after treatment of the subject for an inflammatory disease or condition or an infection. 
     
     
         40 . The method of any one of  claims 34  to  39 , wherein the inflammatory disease or condition is selected from the group consisting of sepsis, septic shock, sterile sepsis, trauma, injury, stroke, acute inflammatory disease, SIRS, acute lung injury, ARDS, pneumonia, necrotizing enterocolitis, acute pancreatitis, renal disease, acute kidney injury, liver injury, acute circulatory failure, preeclampsia, cancer, cancer metastasis, tumor invasion, peripheral artery disease, type 1 or type 2 diabetes, atherosclerotic cardiovascular disease, intermittent claudication, critical limb ischemic disease, myocardial infarction, carotid occlusion, umbilical cord occlusion, low birth-weight, premature birth, surgery-induced inflammation, abscess-induced inflammation, multiple sclerosis, pulmonary insufficiency, peripheral neuropathy, hypoxic ischemia, bacterial infection, wounds, burns, lacerations, contusions, bone fractures, surgical procedures, tissue ischemia, rheumatoid arthritis, meningitis, inflammatory bowel disease, chronic obstructive pulmonary disease, rhinitis, preterm labor, or an infectious disease. 
     
     
         41 . The method of any one of  claims 34  to  39 , wherein the infection is caused by a gram negative bacteria, such as  Neisseria  species including  Neisseria gonorrhoeae  and  Neisseria meningitidis, Branhamella  species including  Branhamella catarrhalis, Escherichia  species including  Escherichia coli, Enterobacter species, Proteus  species including  Proteus mirabilis, Pseudomonas  species including  Pseudomonas aeruginosa, Pseudomonas mallei , and  Pseudomonas pseudomallei, Klebsiella  species including  Klebsiella pneumoniae, Salmonella species, Shigella species, Serratia species, Acinetobacter  species;  Haemophilus  species including  Haemophilus influenzae  and  Haemophilus ducreyi, Brucella species, Yersinia  species including  Yersinia pestis  and  Yersinia enterocolitica, Francisella  species including  Francisella tularensis, Pasteurella  species including  Pasteurella multocida, Vibrio cholerae, Flavobacterium species, meningosepticum, Campylobacter  species including  Campylobacter jejuni, Bacteroides  species (oral, pharyngeal) including  Bacteroides fragilis, Fusobacterium  species including  Fusobacterium nucleatum, Calymmatobacterium granulomatis, Streptobacillus  species including  Streptobacillus moniliformis , and  Legionella  species including  Legionella pneumophila.    
     
     
         42 . The method of any one of  claims 1  to  41 , wherein the subject is a neonate, a child, an adolescent, or an adult. 
     
     
         43 . The method of any one of  claims 1  to  42 , wherein the method is performed one or more times per year. 
     
     
         44 . The method of  claim 43 , wherein the method is performed one or more times per month. 
     
     
         45 . The method of  claim 44 , wherein the method is performed one or more times per week. 
     
     
         46 . The method of  claim 45 , wherein the method is performed one or more times per day. 
     
     
         47 . The method of  claim 46 , wherein the method is performed one or more times per hour. 
     
     
         48 . The method of any one of  claims 1  to  47 , wherein the method is performed at least once, at least twice, at least three times, at least five times, or at least ten times. 
     
     
         49 . The method of any one of  claims 1  to  48 , wherein the method further comprises administering a treatment comprising IAIP or a therapeutic agent to the subject. 
     
     
         50 . The method of  claim 49 , wherein the subject has an IAIP concentration of 200 μg/mL or lower. 
     
     
         51 . The method of  claim 49 , wherein the sample from the subject has an elevated level of IAIP-IAIP ligand complex relative to a reference sample. 
     
     
         52 . The method of any one of  claims 49  to  51 , wherein the subject has or is at risk of developing an inflammatory disease or condition or an infection. 
     
     
         53 . The method of any one of  claims 49  to  52 , wherein the method comprises administering IAIP and a therapeutic agent to the subject. 
     
     
         54 . The method of any one of  claims 49 - 53 , wherein the therapeutic agent is selected from the group consisting of an antibiotic agent, an antiviral agent, an antifungal agent, an antiparasitic agent, an anti-inflammatory agent, an anti-cancer agent, an anti-coagulant, an immunomodulatory agent, a bronchodilator agent, a complement inhibitor, a vasopressor, a sedative, or mechanical ventilation. 
     
     
         55 . The method of any one of  claims 1  to  54 , wherein the subject has been ill for at least one day. 
     
     
         56 . The method of  claim 55 , wherein the subject has been ill for at least one week. 
     
     
         57 . The method of  claim 56 , wherein the subject has been ill for at least one month. 
     
     
         58 . The method of  claim 57 , wherein the subject has been ill for at least one year. 
     
     
         59 . The method of any one of  claims 1  to  58 , wherein the method is for:
 a) evaluating the health status of the subject; 
 b) monitoring the health status of the subject; 
 c) diagnosing the subject as having or being at risk for an inflammatory disease or condition or an infection; 
 d) evaluating efficacy of a treatment administered to the subject; or 
 e) evaluating disease severity in the subject. 
 
     
     
         60 . The method of  claim 59 , wherein the method further comprises comparing the amount of IAIP and/or an IAIP-IAIP ligand complex detected in the sample to the amount of IAIP and/or an IAIP-IAIP ligand complex found in a sample from a normal subject or to a cutoff value. 
     
     
         61 . The method of  claim 60 , wherein an amount of IAIP in the sample that is lower than an amount of IAIP in the sample from the normal subject or relative to the cutoff value indicates that the subject has or is at risk of developing an inflammatory disease or condition or an infection. 
     
     
         62 . The method of  claim 60 , wherein an amount of an IAIP-IAIP ligand complex in the sample that is greater than an amount of IAIP-IAIP ligand complex in the sample from the normal subject or relative to the cutoff value indicates that the subject has or is at risk of developing an inflammatory disease or condition or an infection. 
     
     
         63 . The method of any one of  claims 60  to  62 , wherein the amount of IAIP in the sample from the normal subject, or the cutoff value, is >250 μg/mL. 
     
     
         64 . The method of  claim 63 , wherein the amount of IAIP in the sample from the normal subject is about 260 to about 540 μg/mL. 
     
     
         65 . The method of any one of  claims 1  to  64 , wherein a determination that the subject has an IAIP concentration of 250 μg/mL or less indicates that the subject has or is at high risk of developing an inflammatory disease or condition or an infection or is diagnosed as having an increased risk of morbidity and/or mortality. 
     
     
         66 . The method of any one of  claims 60  to  65 , wherein the subject has an IAIP concentration of 200 to 300 μg/mL and wherein the method is performed at least once a year. 
     
     
         67 . The method of  claim 66 , wherein the method is performed at least twice a year. 
     
     
         68 . The method of  claim 66 , wherein the method is performed at least once a month. 
     
     
         69 . The method of  claim 66 , wherein the method is performed at least once a week. 
     
     
         70 . The method of  claim 66 , wherein the method is performed at least once a day. 
     
     
         71 . The method of  claim 66 , wherein the method is performed at least once an hour. 
     
     
         72 . The method of any one of  claims 1  to  71 , wherein the subject previously had an inflammatory disease or condition or an infection. 
     
     
         73 . The method of any one of  claims 49  to  54 , wherein the method is performed prior to the treatment and one or more times during the course of the treatment. 
     
     
         74 . The method of any one of  claims 49  to  73 , wherein the method is performed after initiation of the treatment and/or after conclusion of the treatment. 
     
     
         75 . The method of any one of  claims 49  to  74 , wherein the treatment is determined to be effective if the concentration of IAIP increases in the subject relative to a prior measurement of IAIP in the subject and/or if the concentration of an IAIP-IAIP ligand complex decreases in the subject relative to a prior measurement of an IAIP-IAIP ligand complex in the subject. 
     
     
         76 . The method of any one of  claims 49  to  74 , wherein the treatment is determined to be ineffective if the concentration of IAIP decreases or remains constant in the subject relative to a prior measurement of IAIP in the subject and/or if the concentration of an IAIP-IAIP ligand complex increases or remains constant in the subject relative to a prior measurement of an IAIP-IAIP ligand complex in the subject. 
     
     
         77 . The method of  claim 76 , wherein the method further comprises modifying or changing the treatment. 
     
     
         78 . A method of treating a subject that has or is at risk of developing an inflammatory disease or infection, wherein the subject has been determined to be in need of treatment according to the method of any one of  claims 1 - 77 , comprising administering to the subject a therapeutically effective amount of IAIP and/or a therapeutic agent selected from the group consisting of an antibiotic agent, an antiviral agent, an antifungal agent, an antiparasitic agent, an anti-inflammatory agent, an anti-cancer agent, an anti-coagulant, an immunomodulatory agent, a bronchodilator agent, a complement inhibitor, a vasopressor, a sedative, or mechanical ventilation. 
     
     
         79 . The method of  claim 78 , wherein the inflammatory disease or condition is selected from the group consisting of sepsis, septic shock, sterile sepsis, trauma, injury, stroke, acute inflammatory disease, SIRS, acute lung injury, ARDS, pneumonia, necrotizing enterocolitis, acute pancreatitis, renal disease, acute kidney injury, liver injury, acute circulatory failure, surgery-induced inflammation, abscess-induced inflammation, multiple sclerosis, preeclampsia, cancer, cancer metastasis, tumor invasion, peripheral artery disease, type 1 or type 2 diabetes, atherosclerotic cardiovascular disease, intermittent claudication, critical limb ischemic disease, myocardial infarction, carotid occlusion, umbilical cord occlusion, low birth-weight, premature birth, pulmonary insufficiency, peripheral neuropathy, hypoxic ischemia, bacterial infection, wounds, burns, lacerations, contusions, bone fractures, surgical procedures, tissue ischemia, rheumatoid arthritis, meningitis, inflammatory bowel disease, chronic obstructive pulmonary disease, rhinitis, preterm labor, or an infectious disease. 
     
     
         80 . The method of  claim 78 , wherein the infection is caused by a gram negative bacteria, such as  Neisseria  species including  Neisseria gonorrhoeae  and  Neisseria meningitidis, Branhamella  species including  Branhamella catarrhalis, Escherichia  species including  Escherichia coli, Enterobacter  species,  Proteus  species including  Proteus mirabilis, Pseudomonas  species including  Pseudomonas aeruginosa, Pseudomonas mallei , and  Pseudomonas pseudomallei, Klebsiella  species including  Klebsiella pneumoniae, Salmonella species, Shigella species, Serratia species, Acinetobacter  species;  Haemophilus  species including  Haemophilus influenzae  and  Haemophilus ducreyi, Brucella species, Yersinia  species including  Yersinia pestis  and  Yersinia enterocolitica, Francisella  species including  Francisella tularensis, Pasteurella  species including  Pasteurella multocida, Vibrio cholerae, Flavobacterium species, meningosepticum, Campylobacter  species including  Campylobacter jejuni, Bacteroides  species (oral, pharyngeal) including  Bacteroides fragilis, Fusobacterium  species including  Fusobacterium nucleatum, Calymmatobacterium granulomatis, Streptobacillus  species including  Streptobacillus moniliformis , and  Legionella  species including  Legionella pneumophila.    
     
     
         81 . A kit for quantifying IAIP or an IAIP-IAIP ligand complex in a sample, wherein the kit comprises an IAIP binding agent and an IAIP detection agent and, optionally, one or more of the following: a wash buffer, a blocking agent, a substrate for detection of a label, and instructions for quantifying a level of IAIP or an IAIP-IAIP ligand complex in a sample. 
     
     
         82 . The kit of  claim 81 , wherein the binding agent is immobilized on a support. 
     
     
         83 . The kit of  claim 81  or  82 , wherein the detection agent is labeled. 
     
     
         84 . The kit of any one of  claims 81  to  83 , wherein the IAIP binding agent is an IAIP-specific antibody or an IAIP ligand. 
     
     
         85 . The kit of any one of  claims 81  to  84 , wherein the kit further comprises an IAIP ligand binding agent. 
     
     
         86 . The kit of  claim 85 , wherein the IAIP ligand binding agent is an antibody that binds to an IAIP ligand. 
     
     
         87 . The kit of any one of  claims 81  to  86 , wherein the IAIP detection agent is an IAIP-specific antibody or an IAIP ligand. 
     
     
         88 . The kit of any one of  claims 81  to  87 , wherein the kit further comprises an IAIP ligand detection agent. 
     
     
         89 . The kit of  claim 88 , wherein the IAIP ligand detection agent is an antibody that binds specifically to an IAIP ligand. 
     
     
         90 . The kit of  claim 84  or  87 , wherein the IAIP-specific antibody is a monoclonal antibody. 
     
     
         91 . The kit of  claim 90 , wherein the monoclonal antibody is MAb 69.26 or MAb 69.31. 
     
     
         92 . The kit of any one of  claims 82  to  91 , wherein the support is a plate, a resin, a container, a membrane, a biochip, a particle, a nanoparticle, a test strip, or a bead. 
     
     
         93 . The kit of any one of  claims 81  to  92 , wherein the label is an enzyme, an enzyme substrate, biotin, a particle, a fluorescent dye, a luminescent compound, or a radiolabel. 
     
     
         94 . The kit of any one of  claims 84 ,  86 ,  87 ,  88 , and  89 , wherein the IAIP ligand is selected from the group consisting of endotoxin (LPS), heparin, a histone, hyaluronic acid, laminin, tenascin C, aggrecan, vitronectin, fibronectin, von Willebrand Factor, pentraxin-3 (PTX3), TNF-stimulated gene-6 (TSG-6), factor IX, a complement component, factor XIIIa, and tissue transglutaminase. 
     
     
         95 . The method of  claim 1 , wherein the binding agent is an IAIP ligand that binds to IAIP. 
     
     
         96 . The method of  claim 1 , wherein the binding agent is an antibody that specifically binds to IAIP. 
     
     
         97 . The method of  claim 1 ,  95  or  96 , wherein the detection agent comprises an IAIP ligand. 
     
     
         98 . The method of  claim 97 , wherein the detection agent further comprises an antibody that binds to the IAIP ligand detection agent. 
     
     
         99 . The method of  claim 1 ,  95 , or  96 , wherein the detection agent is an antibody that specifically binds to IAIP. 
     
     
         100 . The method of  claim 8  or  11 , wherein the detection agent comprises an IAIP ligand that binds to IAIP. 
     
     
         101 . The method of  claim 100 , wherein the detection agent further comprises an antibody that binds to the IAIP ligand detection agent. 
     
     
         102 . The method of  claim 100 , wherein the IAIP ligand of the IAIP-IAIP ligand complex is different from the IAIP ligand detection agent. 
     
     
         103 . The method of  claim 8 ,  9 , or  12 , wherein the detection agent is an antibody that binds to the IAIP ligand of the IAIP-IAIP ligand complex. 
     
     
         104 . The method of  claim 8  or  11 , wherein the detection agent is an antibody that specifically binds to IAIP of the IAIP-IAIP ligand complex. 
     
     
         105 . The method of  claim 96 , wherein the antibody is a monoclonal antibody. 
     
     
         106 . The method of  claim 97 , wherein the antibody is a monoclonal antibody. 
     
     
         107 . The method of  claim 99 , wherein the antibody is a monoclonal antibody. 
     
     
         108 . The method of  claim 101 , wherein the antibody is a monoclonal antibody. 
     
     
         109 . The method of  claim 103 , wherein the antibody is a monoclonal antibody. 
     
     
         110 . The method of  claim 104 , wherein the antibody is a monoclonal antibody. 
     
     
         111 . The method of  claim 96 , wherein the antibody is MAb 69.26 or MAb 69.31. 
     
     
         112 . The method of  claim 99 , wherein the antibody is MAb 69.26 or MAb 69.31 
     
     
         113 . The method of  claim 104 , wherein the antibody is MAb 69.26 or MAb 69.31 
     
     
         114 . The method of  claim 95 , wherein the IAIP ligand is selected from the group consisting of endotoxin (LPS), heparin, a histone, hyaluronic acid, vitronectin, fibronectin, laminin, tenascin C, aggrecan, von Willebrand Factor, pentraxin-3 (PTX3), TNF-stimulated gene-6 (TSG-6), factor IX, a complement component, factor XIIIa, and tissue transglutaminase. 
     
     
         115 . The method of  claim 97 , wherein the IAIP ligand is selected from the group consisting of endotoxin (LPS), heparin, a histone, hyaluronic acid, vitronectin, fibronectin, laminin, tenascin C, aggrecan, von Willebrand Factor, pentraxin-3 (PTX3), TNF-stimulated gene-6 (TSG-6), factor IX, a complement component, factor XIIIa, and tissue transglutaminase. 
     
     
         116 . The method of  claim 100 , wherein the IAIP ligand is selected from the group consisting of endotoxin (LPS), heparin, a histone, hyaluronic acid, vitronectin, fibronectin, laminin, tenascin C, aggrecan, von Willebrand Factor, pentraxin-3 (PTX3), TNF-stimulated gene-6 (TSG-6), factor IX, a complement component, factor XIIIa, and tissue transglutaminase. 
     
     
         117 . The method of  claim 8 , wherein the IAIP ligand is selected from the group consisting of endotoxin (LPS), heparin, a histone, hyaluronic acid, vitronectin, fibronectin, laminin, tenascin C, aggrecan, von Willebrand Factor, pentraxin-3 (PTX3), TNF-stimulated gene-6 (TSG-6), factor IX, a complement component, factor XIIIa, and tissue transglutaminase. 
     
     
         118 . The method of any one of  claims 114 - 117 , wherein the complement component is C1q, C2, C3, C4, C5, C6, C8, properdin, or factor D. 
     
     
         119 . The method of  claim 1 , wherein the detection agent comprises a label. 
     
     
         120 . The method of  claim 119 , wherein the label is biotin, an enzyme, an enzyme substrate, a radiolabel, a luminescent compound, colloidal gold, a particle, or a fluorescent dye. 
     
     
         121 . The method of  claim 1 , wherein the support is a plate, a particle, a nanoparticle, a resin, a membrane, a biochip, a container, a test strip, or a bead. 
     
     
         122 . The method of  claim 1 , wherein the method further includes a wash step between steps a) and b). 
     
     
         123 . The method of  claim 1 , wherein the method further includes a wash step between steps b) and c). 
     
     
         124 . The method of  claim 1 , wherein the method further includes a blocking step prior to step a) or step b). 
     
     
         125 . The method of  claim 1 , wherein the contacting in step a) and/or b) is performed at a pH of about 7.0 to about 3.5. 
     
     
         126 . The method of  claim 125 , wherein the pH is about 5.0 to about 3.5. 
     
     
         127 . The method of  claim 126 , wherein the pH is about 4.0. 
     
     
         128 . The method of  claim 1 , wherein the sample is a fluid. 
     
     
         129 . The method of  claim 128 , wherein the fluid is blood, plasma, serum, urine, cerebrospinal fluid, synovial fluid, amniotic fluid, interstitial fluid, follicular fluid, peritoneal fluid, bronchoalveolar lavage fluid, breast milk, sputum, lymph, bile, or tissue homogenate. 
     
     
         130 . The method of  claim 1 , wherein the subject is a human subject. 
     
     
         131 . The method of  claim 130 , wherein the subject has been identified as having or at risk of developing an inflammatory disease or condition or an infection. 
     
     
         132 . The method of  claim 130 , wherein the subject has not been identified as having or at risk of developing an inflammatory disease or condition or an infection. 
     
     
         133 . The method of  claim 130 , wherein the method is performed before, after, or concurrent with diagnosis of the subject as having or at risk of an inflammatory disease or condition or an infection. 
     
     
         134 . The method of  claim 130 , wherein the method is performed substantially concurrent with treatment of the subject for an inflammatory disease or condition or an infection. 
     
     
         135 . The method of  claim 130 , wherein the method is performed prior to treatment of the subject for an inflammatory disease or condition or an infection. 
     
     
         136 . The method of  claim 130 , wherein the method is performed after treatment of the subject for an inflammatory disease or condition or an infection. 
     
     
         137 . The method of  claim 131 , wherein the inflammatory disease or condition is selected from the group consisting of sepsis, septic shock, sterile sepsis, trauma, injury, stroke, acute inflammatory disease, SIRS, acute lung injury, ARDS, pneumonia, necrotizing enterocolitis, acute pancreatitis, renal disease, acute kidney injury, liver injury, acute circulatory failure, preeclampsia, cancer, cancer metastasis, tumor invasion, peripheral artery disease, type 1 or type 2 diabetes, atherosclerotic cardiovascular disease, intermittent claudication, critical limb ischemic disease, myocardial infarction, carotid occlusion, umbilical cord occlusion, low birth-weight, premature birth, surgery-induced inflammation, abscess-induced inflammation, multiple sclerosis, pulmonary insufficiency, peripheral neuropathy, hypoxic ischemia, bacterial infection, wounds, burns, lacerations, contusions, bone fractures, surgical procedures, tissue ischemia, rheumatoid arthritis, meningitis, inflammatory bowel disease, chronic obstructive pulmonary disease, rhinitis, preterm labor, or an infectious disease. 
     
     
         138 . The method of  claim 131 , wherein the infection is caused by a gram negative bacteria, such as  Neisseria  species including  Neisseria gonorrhoeae  and  Neisseria meningitidis, Branhamella  species including  Branhamella catarrhalis, Escherichia  species including  Escherichia coli, Enterobacter  species,  Proteus  species including  Proteus mirabilis, Pseudomonas  species including  Pseudomonas aeruginosa, Pseudomonas mallei , and  Pseudomonas pseudomallei, Klebsiella  species including  Klebsiella pneumoniae, Salmonella species, Shigella species, Serratia species, Acinetobacter  species;  Haemophilus  species including  Haemophilus influenzae  and  Haemophilus ducreyi, Brucella  species,  Yersinia  species including  Yersinia pestis  and  Yersinia enterocolitica, Francisella  species including  Francisella tularensis, Pasturella  species including  Pasteurella multocida, Vibrio cholerae, Flavobacterium species, meningosepticum, Campylobacter  species including  Campylobacter jejuni, Bacteroides  species (oral, pharyngeal) including  Bacteroides fragilis, Fusobacterium  species including  Fusobacterium nucleatum, Calymmatobacterium granulomatis, Streptobacillus  species including  Streptobacillus moniliformis , and  Legionella  species including  Legionella pneumophila.    
     
     
         139 . The method of  claim 1 , wherein the subject is a neonate, a child, an adolescent, or an adult. 
     
     
         140 . The method of  claim 1 , wherein the method is performed one or more times per year. 
     
     
         141 . The method of  claim 140 , wherein the method is performed one or more times per month. 
     
     
         142 . The method of  claim 141 , wherein the method is performed one or more times per week. 
     
     
         143 . The method of  claim 142 , wherein the method is performed one or more times per day. 
     
     
         144 . The method of  claim 143 , wherein the method is performed one or more times per hour. 
     
     
         145 . The method of  claim 1 , wherein the method is performed at least once, at least twice, at least three times, at least five times, or at least ten times. 
     
     
         146 . The method of  claim 1 , wherein the method further comprises administering a treatment comprising IAIP or a therapeutic agent to the subject. 
     
     
         147 . The method of  claim 146 , wherein the subject has an IAIP concentration of 200 μg/mL or lower. 
     
     
         148 . The method of  claim 146 , wherein the sample from the subject has an elevated level of IAIP-IAIP ligand complex relative to a reference sample. 
     
     
         149 . The method of  claim 146 , wherein the subject has or is at risk of developing an inflammatory disease or condition or an infection. 
     
     
         150 . The method of  claim 146 , wherein the method comprises administering IAIP and a therapeutic agent to the subject. 
     
     
         151 . The method of  claim 146  or  150 , wherein the therapeutic agent is selected from the group consisting of an antibiotic agent, an antiviral agent, an antifungal agent, an antiparasitic agent, an anti-inflammatory agent, an anti-cancer agent, an anti-coagulant, an immunomodulatory agent, a bronchodilator agent, a complement inhibitor, a vasopressor, a sedative, or mechanical ventilation. 
     
     
         152 . The method of  claim 1 , wherein the subject has been ill for at least one day. 
     
     
         153 . The method of  claim 152 , wherein the subject has been ill for at least one week. 
     
     
         154 . The method of  claim 153 , wherein the subject has been ill for at least one month. 
     
     
         155 . The method of  claim 154 , wherein the subject has been ill for at least one year. 
     
     
         156 . The method of  claim 1 , wherein the method is for:
 c) evaluating the health status of the subject;   d) monitoring the health status of the subject;   c) diagnosing the subject as having or being at risk for an inflammatory disease or condition or an infection;   d) evaluating efficacy of a treatment administered to the subject; or   e) evaluating disease severity in the subject.   
     
     
         157 . The method of  claim 156 , wherein the method further comprises comparing the amount of IAIP and/or an IAIP-IAIP ligand complex detected in the sample to the amount of IAIP and/or an IAIP-IAIP ligand complex found in a sample from a normal subject or to a cutoff value. 
     
     
         158 . The method of  claim 157 , wherein an amount of IAIP in the sample that is lower than an amount of IAIP in the sample from the normal subject or relative to the cutoff value indicates that the subject has or is at risk of developing an inflammatory disease or condition or an infection. 
     
     
         159 . The method of  claim 157 , wherein an amount of an IAIP-IAIP ligand complex in the sample that is greater than an amount of IAIP-IAIP ligand complex in the sample from the normal subject or relative to the cutoff value indicates that the subject has or is at risk of developing an inflammatory disease or condition or an infection. 
     
     
         160 . The method of  claim 157 , wherein the amount of IAIP in the sample from the normal subject, or the cutoff value, is >250 μg/mL. 
     
     
         161 . The method of  claim 160 , wherein the amount of IAIP in the sample from the normal subject is about 260 to about 540 μg/mL. 
     
     
         162 . The method of  claim 1 , wherein a determination that the subject has an IAIP concentration of 250 μg/mL or less indicates that the subject has or is at high risk of developing an inflammatory disease or condition or an infection or is diagnosed as having an increased risk of morbidity and/or mortality. 
     
     
         163 . The method of  claim 157 , wherein the subject has an IAIP concentration of 200 to 300 μg/mL and wherein the method is performed at least once a year. 
     
     
         164 . The method of  claim 163 , wherein the method is performed at least twice a year. 
     
     
         165 . The method of  claim 163 , wherein the method is performed at least once a month. 
     
     
         166 . The method of  claim 163 , wherein the method is performed at least once a week. 
     
     
         167 . The method of  claim 163 , wherein the method is performed at least once a day. 
     
     
         168 . The method of  claim 163 , wherein the method is performed at least once an hour. 
     
     
         169 . The method of  claim 1 , wherein the subject previously had an inflammatory disease or condition or an infection. 
     
     
         170 . The method of  claim 146 , wherein the method is performed prior to the treatment and one or more times during the course of the treatment. 
     
     
         171 . The method of  claim 146 , wherein the method is performed after initiation of the treatment and/or after conclusion of the treatment. 
     
     
         172 . The method of  claim 146  wherein the treatment is determined to be effective if the concentration of IAIP increases in the subject relative to a prior measurement of IAIP in the subject and/or if the concentration of an IAIP-IAIP ligand complex decreases in the subject relative to a prior measurement of an IAIP-IAIP ligand complex in the subject. 
     
     
         173 . The method of  claim 146 , wherein the treatment is determined to be ineffective if the concentration of IAIP decreases or remains constant in the subject relative to a prior measurement of IAIP in the subject and/or if the concentration of an IAIP-IAIP ligand complex increases or remains constant in the subject relative to a prior measurement of an IAIP-IAIP ligand complex in the subject. 
     
     
         174 . The method of  claim 173 , wherein the method further comprises modifying or changing the treatment. 
     
     
         175 . A method of treating a subject that has or is at risk of developing an inflammatory disease or infection, wherein the subject has been determined to be in need of treatment according to the method of  claim 1 , comprising administering to the subject a therapeutically effective amount of IAIP and/or a therapeutic agent selected from the group consisting of an antibiotic agent, an antiviral agent, an antifungal agent, an antiparasitic agent, an anti-inflammatory agent, an anti-cancer agent, an anti-coagulant, an immunomodulatory agent, a bronchodilator agent, a complement inhibitor, a vasopressor, a sedative, or mechanical ventilation. 
     
     
         176 . The method of  claim 175 , wherein the inflammatory disease or condition is selected from the group consisting of sepsis, septic shock, sterile sepsis, trauma, injury, stroke, acute inflammatory disease, SIRS, acute lung injury, ARDS, pneumonia, necrotizing enterocolitis, acute pancreatitis, renal disease, acute kidney injury, liver injury, acute circulatory failure, surgery-induced inflammation, abscess-induced inflammation, multiple sclerosis, preeclampsia, cancer, cancer metastasis, tumor invasion, peripheral artery disease, type 1 or type 2 diabetes, atherosclerotic cardiovascular disease, intermittent claudication, critical limb ischemic disease, myocardial infarction, carotid occlusion, umbilical cord occlusion, low birth-weight, premature birth, pulmonary insufficiency, peripheral neuropathy, hypoxic ischemia, bacterial infection, wounds, burns, lacerations, contusions, bone fractures, surgical procedures, tissue ischemia, rheumatoid arthritis, meningitis, inflammatory bowel disease, chronic obstructive pulmonary disease, rhinitis, preterm labor, or an infectious disease. 
     
     
         177 . The method of  claim 175 , wherein the infection is caused by a gram negative bacteria, such as  Neisseria  species including  Neisseria gonorrhoeae  and  Neisseria meningitidis, Branhamella  species including  Branhamella catarrhalis, Escherichia  species including  Escherichia coli, Enterobacter  species,  Proteus  species including  Proteus mirabilis, Pseudomonas  species including  Pseudomonas aeruginosa, Pseudomonas mallei , and  Pseudomonas pseudomallei, Klebsiella  species including  Klebsiella pneumoniae, Salmonella species, Shigella species, Serratia species, Acinetobacter  species;  Haemophilus  species including  Haemophilus influenzae  and  Haemophilus ducreyi, Brucella  species,  Yersinia  species including  Yersinia pestis  and  Yersinia enterocolitica, Francisella  species including  Francisella tularensis, Pasturella  species including  Pasteurella multocida, Vibrio cholerae, Flavobacterium species, meningosepticum, Campylobacter  species including  Campylobacter jejuni, Bacteroides  species (oral, pharyngeal) including  Bacteroides fragilis, Fusobacterium  species including  Fusobacterium nucleatum, Calymmatobacterium granulomatis, Streptobacillus  species including  Streptobacillus moniliformis , and  Legionella  species including  Legionella pneumophila.    
     
     
         178 . The kit of  claim 81 , wherein the detection agent is labeled. 
     
     
         179 . The kit of  claim 81 , wherein the IAIP binding agent is an IAIP-specific antibody or an IAIP ligand. 
     
     
         180 . The kit of  claim 81 , wherein the kit further comprises an IAIP ligand binding agent. 
     
     
         181 . The kit of  claim 180 , wherein the IAIP ligand binding agent is an antibody that binds to an IAIP ligand. 
     
     
         182 . The kit of  claim 81 , wherein the IAIP detection agent is an IAIP-specific antibody or an IAIP ligand. 
     
     
         183 . The kit of  claim 81 , wherein the kit further comprises an IAIP ligand detection agent. 
     
     
         184 . The kit of  claim 183 , wherein the IAIP ligand detection agent is an antibody that binds specifically to an IAIP ligand. 
     
     
         185 . The kit of  claim 179  or  182 , wherein the IAIP-specific antibody is a monoclonal antibody. 
     
     
         186 . The kit of  claim 185 , wherein the monoclonal antibody is MAb 69.26 or MAb 69.31. 
     
     
         187 . The kit of  claim 82 , wherein the support is a plate, a resin, a container, a membrane, a biochip, a particle, a nanoparticle, a test strip, or a bead. 
     
     
         188 . The kit of  claim 81 , wherein the label is an enzyme, an enzyme substrate, biotin, a particle, a fluorescent dye, a luminescent compound, or a radiolabel. 
     
     
         189 . The kit of any one of  claims 179 - 184 , wherein the IAIP ligand is selected from the group consisting of endotoxin (LPS), heparin, a histone, hyaluronic acid, laminin, tenascin C, aggrecan, vitronectin, fibronectin, von Willebrand Factor, pentraxin-3 (PTX3), TNF-stimulated gene-6 (TSG-6), factor IX, a complement component, factor XIIIa, and tissue transglutaminase.