Monoclonal and oligoclonal anti-egfr antibodies for use in the treatment of tumors expressing predominantly high affinity egfr ligands or tumors expressing predominantly low affinity egfr ligands

Abstract

Disclosed are pharmaceutical preparations for, and methods for determining, appropriate and effective treatment with therapeutic agents comprising a single species of anti-EGFR monoclonal antibody or therapeutic agents comprising a plurality of species of such antibodies, as well as kits useful for making such determinations.

What is claimed is: 
     
         1 . A kit for testing a tumor sample to determine levels of both high and low affinity EGFR ligands in the sample, said kit comprising a container comprising; a) at least two pairs of high affinity EGFR ligand-specific polymerase chain reaction (PCR) primers, b) at least two pairs of low affinity EGFR ligand-specific PCR primers, and c) at least one reverse transcription PCR (RT-PCR) reagent. 
     
     
         2 . The kit of  claim 1 , wherein the at least two pairs of high affinity EGFR ligand-specific polymerase chain reaction primers are specific to at least two of betacellulin, EGF, HB-EGF or TGF alpha and each of the at least two pairs of low affinity EGFR ligand-specific polymerase chain reaction primers are specific to at least two of amphiregulin, epigen, or epiregulin. 
     
     
         3 . The kit of  claim 2 , wherein the at least two pairs of high affinity EGFR ligand-specific primers comprise specific primer pairs for betacellulin, EGF, HB-EGF and TGF alpha and the at least two pairs of high affinity EGFR ligand-specific primers comprise specific primer pairs for all of amphiregulin, epigen, and epiregulin. 
     
     
         4 . The kit of  claim 2 , further comprising at least one fluorescent reporter molecule. 
     
     
         5 . The kit of  claim 1 , wherein the at least one RT-PCR reagent is one or more of an RNA-dependent DNA polymerase, a DNA-dependent DNA polymerase, or a solution comprising at least a micromolar concentration of each of ATP, GTP, CTP and TTP. 
     
     
         6 . The kit of  claim 2 , wherein the at least one RT-PCR reagent is one or more of an RNA-dependent DNA polymerase, a DNA-dependent DNA polymerase, or a solution comprising at least a micromolar concentration of each of ATP, GTP, CTP and TTP. 
     
     
         7 . The kit of  claim 5  wherein the primers, the reagents, or both, have been prepared under current Good Manufacturing Practice (cGMP) conditions. 
     
     
         8 . The kit of  claim 5 , wherein container further comprises total RNA isolated from a tumor sample. 
     
     
         9 . The kit of  claim 8 , wherein the tumor sample from which the RNA is isolated is a fresh, fixed, frozen, or formalin fixed paraffin embedded (FFPE) sample. 
     
     
         10 . The kit of  claim 9 , wherein the tumor sample is a circulating tumor cell sample. 
     
     
         11 . The kit of  claim 9 , wherein the tumor sample is a biopsy sample. 
     
     
         12 . The kit of  claim 9 , wherein the tumor sample is from a tumor of the skin, central nervous system, head, neck, esophagus, stomach, colon, rectum, anus, liver, pancreas, bile duct, gallbladder, lung, breast, ovary, uterus, cervix, vagina, testis, germ cells, prostate, kidney, ureter, urinary bladder, adrenal, pituitary, thyroid, bone, muscle or connective tissue. 
     
     
         13 . A kit for testing a tumor sample to determine levels of both high and low affinity EGFR ligands in the sample, said kit comprising one or more containers, one of said containers comprising; a) at least two pairs of high affinity EGFR ligand-specific polymerase chain reaction (PCR) primers, b) at least two pairs of low affinity EGFR ligand-specific PCR primers, c) at least one reverse transcription PCR (RT-PCR) reagent, and d) RNA isolated from a tumor sample. 
     
     
         14 . The kit of  claim 13 , wherein the container comprising a), b), c), and d) is a 96 well plate. 
     
     
         15 . The kit of  claim 13 , wherein the tumor sample from which the RNA is isolated is a fresh, fixed, frozen, or formalin fixed paraffin embedded (FFPE) sample. 
     
     
         16 . The kit of  claim 15 , wherein the tumor sample is a circulating tumor cell sample. 
     
     
         17 . The kit of  claim 16 , wherein the tumor sample is a biopsy sample. 
     
     
         18 . The kit of  claim 15 , wherein the tumor sample was from a tumor of the skin, central nervous system, head, neck, esophagus, stomach, colon, rectum, anus, liver, pancreas, bile duct, gallbladder, lung, breast, ovary, uterus, cervix, vagina, testis, germ cells, prostate, kidney, ureter, urinary bladder, adrenal, pituitary, thyroid, bone, muscle or connective tissue. 
     
     
         19 . The kit of  claim 15 , wherein the container comprising a), b), c), and d) is a 96 well plate 
     
     
         20 . The kit of  claim 18 , wherein the container comprising a), b), c), and d) is a 96 well plate. 
     
     
         21 . A monoclonal anti-EGFR antibody preparation comprising a single species of monoclonal antibody for treatment of a patient predicted by a method to have a favorable outcome from treatment with the monoclonal anti-EGFR antibody preparation by determining whether a patient having a tumor is predicted to have a favorable outcome, comprising reduction of growth of the tumor, as a result of treatment #1 with a monoclonal anti-EGFR antibody preparation comprising a single species of monoclonal antibody, and as a result of treatment #2 with an oligoclonal anti-EGFR antibody preparation comprising a plurality of species of monoclonal anti-EGFR antibodies, one against each of at least two extracellular epitopes of EGFR,
 or,   
       whether the patient is predicted to have the favorable outcome as a result of treatment #2 but not as a result of treatment #1;
 the method comprising: 
 
       obtaining a biopsy sample of the tumor and:
 a) measuring levels of protein or mRNAs coding for at least two low affinity EGFR ligands selected from amphiregulin, epigen, or epiregulin in the biopsy sample, 
 b) measuring levels of protein or mRNAs coding for at least two high affinity EGFR ligands selected from betacellulin, EGF, HB-EGF or TGFα in the biopsy sample, and 
 c) comparing the average level of protein or mRNAs coding for each of the high affinity EGFR ligands measured in a) to the average level of protein or mRNAs coding for each of the low affinity EGFR ligands measured in b); 
 wherein, 
 
       if the average level of protein or mRNAs coding for low affinity EGFR ligands measured in a) is greater than the average level of protein or mRNAs coding for high affinity EGFR ligands measured in b), the patient is predicted to have the favorable outcome as a result of treatment #1 and the patient is also predicted to have the favorable outcome as a result of treatment #2, and 
       if the average level of protein or mRNAs coding for low affinity EGFR ligands measured in a) is less than or equal to the average level of protein or mRNAs coding for high affinity EGFR ligands measured in b), the patient is predicted to have an unfavorable outcome from treatment #1 and is predicted to have a favorable outcome from treatment #2.