Vaccine-induced hepatitis B viral strain and uses thereof

Abstract

This invention provides an isolated strain of Hepatitis B virus designated Human Hepatitis B Virus Surface Antigen-‘S’-145 Singapore Strain (Glycine to Arginine) which constituent viral genome is deposited under Accession Nos. P97121504, P97121505 and P97121506 with the European Collection of Cell Culture on 15 th Dec. 1997. This invention also provides an isolated nucleic acid encoding a polypeptide which is a mutant major surface antigen of a strain of hepatitis B virus, such polypeptide having an amino acid sequence which differs from the amino acid sequence of a major surface antigen of a wild type hepatitis B virus in that the amino acid at position number 145 of such polypeptide is an arginine rather than a glycine, and the purified peptide This invention further provides an isolated nucleic acid which encodes a peptide, wherein the peptide is encoded by a nucleic acid molecule comprising nucleotides 527 through 595 of SEQ. I.D. No. 1, and the purified peptide. This invention also provides various methods using the disclosed isolated nucleic acids and polypeptides.

1 - 11 . (canceled)  
     
     
         12 . A vector system for the production of a polypeptide which comprises, in a suitable host, a vector comprising an isolated nucleic acid encoding a polypeptide which is a mutant major surface antigen of a strain of hepatitis B virus, such polypeptide having an amino acid sequence which differs from the amino acid sequence of a major surface antigen of a wild type hepatitis B virus in that the amino acid at position number 145 of such polypeptide is an arginine rather than a glycine and operatively linked to a promoter of RNA transcription.  
     
     
         13 . A vector system for the production of a polypeptide which comprises, in a suitable host, a vector comprising an isolated nucleic acid encoding a peptide, wherein the peptide is encoded by a nucleic acid molecule comprising nucleotides 527 through 595 of SEQ. I.D. No. 1.  
     
     
         14 - 16 . (canceled)  
     
     
         17 . A method of producing a polypeptide which comprises growing the host vector system of  claim 12  under suitable conditions permitting production of the polypeptide and recovering the polypeptide so produced.  
     
     
         18 . A method of producing a peptide which comprises growing the host vector system of  claim 13  under suitable conditions permitting production of the polypeptide and recovering the polypeptide so produced.  
     
     
         19 . A method of obtaining a polypeptide in purified form which comprises: 
 (a) introducing into a suitable host cell a vector comprising an isolated nucleic acid encoding a polypeptide which is a mutant major surface antigen of a strain of hepatitis B virus, such polypeptide having an amino acid sequence which differs from the amino acid sequence of a major surface antigen of a wild type hepatitis B virus in that the amino acid at position number 145 of such polypeptide is an arginine rather than a glycine and operatively linked to a promoter of RNA transcription;    (b) culturing the resulting host cell so as to produce the polypeptide;    (c) recovering the polypeptide produced in step (b); and    (d) purifying the polypeptide so recovered.    
     
     
         20 . A method of obtaining a peptide in purified form which comprises: 
 (a) introducing into a suitable host cell a vector comprising an isolated nucleic acid encoding a peptide, wherein the peptide is encoded by a nucleic acid molecule comprising nucleotides 527 through 595 of SEQ. I.D. No. 1;    (b) culturing the resulting host cell so as to produce the polypeptide;    (c) recovering the polypeptide produced in step (b); and    (d) purifying the polypeptide so recovered.    
     
     
         21 . A purified polypeptide which is a mutant major surface antigen of a strain of hepatitis B virus, such polypeptide having an amino acid sequence which differs from the amino acid sequence of a major surface antigen of a wild type hepatitis B virus in that the amino acid at position number 145 of such polypeptide is an arginine rather than a glycine.  
     
     
         22 . A purified polypeptide obtained from the method of  claim 19 .  
     
     
         23 . A purified peptide, wherein the peptide has an amino acid sequence comprising amino acid residues 298 through 320 of the amino acid sequence designated SEQ. I.D. No. 3.  
     
     
         24 . A purified peptide obtained from the method of  claim 20 .  
     
     
         25 - 26 . (canceled)  
     
     
         27 . A method of obtaining antibodies to a polypeptide which is a mutant major surface antigen of a strain of hepatitis B virus, such polypeptide having an amino acid sequence which differs from the amino acid sequence of a major surface antigen of a wild type hepatitis B virus in that the amino acid at position number 145 of such polypeptide is an arginine, rather than a glycine, and not to the major surface antigen of a wild type hepatitis B virus, comprising: 
 (a) obtaining the polypeptide in a purified form;    (b) immunizing an organism capable of producing antibodies against the purified polypeptide;    (c) collecting the produced antibodies;    (d) combining the produced antibodies and the purified polypeptide under conditions to form a complex; and    (e) determining which produced antibodies form a complex with the purified polypeptide so as to obtain antibodies to the polypeptide.    
     
     
         28 . The method of  claim 27 , wherein the polypeptide is being encoded by nucleotides 155 through 835 of the nucleic acid sequence designated SEQ. I.D. No. 1.  
     
     
         29 . The method of  claim 27 , wherein the polypeptide has an amino acid sequence substantially the same as amino acid residues 174 through 400 of the amino acid sequence designated SEQ. I.D. No. 3.  
     
     
         30 . The method of  claim 27 , wherein the organism comprises a rabbit or a mouse.  
     
     
         31 . A method of obtaining antibodies to a peptide, wherein the peptide has an amino acid sequence comprising amino acid residues 298 through 320 of the amino acid sequence designated SEQ. I.D. No. 3, comprising: 
 (a) obtaining the peptide in a purified form;    (b) immunizing an organism capable of producing antibodies against the purified peptide;    (c) collecting the produced antibodies;    (d) combining the produced antibodies and the purified peptide under conditions to form a complex; and    (e) determining which produced antibodies form a complex with the purified peptide so as to obtain antibodies to the peptide.    
     
     
         32 . The method of  claim 31 , wherein the organism comprises a rabbit or a mouse.  
     
     
         33 - 36 . (canceled)  
     
     
         37 . A method for use of a nucleic acid encoding a polypeptide which is a mutant major surface antigen of a strain of hepatitis B virus, such polypeptide having an amino acid sequence which differs from the amino acid sequence of a major surface antigen of a wild type hepatitis B virus in that the amino acid at position number 145 of such polypeptide is an arginine, rather than a glycine for determining whether a subject is infected with a strain of Hepatitis B virus designated Human Hepatitis B Virus Surface Antigen-‘S’-145 Singapore Strain (Glycine to Arginine), wherein such method comprises 
 (a) obtaining an appropriate nucleic acid sample from the subject; and    (b) determining whether the nucleic acid sample from step (a) is, or is derived from, a nucleic acid encoding a polypeptide which is a mutant major surface antigen of a strain of hepatitis B virus, such polypeptide having an amino acid sequence which differs from the amino acid sequence of a major surface antigen of a wild type hepatitis B virus in that the amino acid at position number 145 of such polypeptide is an arginine, rather than a glycine.    
     
     
         38 . (canceled)  
     
     
         39 . The method of  claim 37 , wherein the nucleic acid sample in step (a) comprises mRNA corresponding to the transcript of DNA encoding a polypeptide which is a mutant major surface antigen of a strain of hepatitis B virus, such polypeptide having an amino acid sequence which differs from the amino acid sequence of a major surface antigen of a wild type hepatitis B virus in that the amino acid at position number 145 of such polypeptide is an arginine, rather than a glycine, and wherein the determining of step (b) comprises: 
 (i translating the mRNA under suitable conditions to obtain an amino acid sequence; and    (ii comparing the amino acid sequence of step (i) with the amino acid sequence encoded by an isolated nucleic acid encoding a polypeptide which is a mutant major surface antigen of a strain of hepatitis B virus, such polypeptide having an amino acid sequence which differs from the amino acid sequence of a major surface antigen of a wild type hepatitis B virus in that the amino acid at position number 145 of such polypeptide is an arginine rather than a glycine, wherein the polypeptide has an amino acid sequence substantially the same as amino acid residues 174 through 400 of the amino acid sequence designated SEO. ID NO. 3 so as to determine whether the nucleic acid sample is, or is derived from, a nucleic acid which encodes the polypeptide.    
     
     
         40 . The method of  claim 37 , wherein the determining of step (b) comprises: 
 (i) amplifying the nucleic acid present in the sample of step (a); and    (ii) detecting the presence of polypeptide in the resulting amplified nucleic acid.    
     
     
         41 . (canceled)  
     
     
         42 . The method of  claim 37 , wherein the isolated nucleic acid, is labeled with a detectable marker.  
     
     
         43 . The method of  claim 42 , wherein the detectable marker is a radioactive isotope, a fluorophor, or an enzyme.  
     
     
         44 . The method of  claim 37 , wherein the sample comprises blood, tissue, or sera.  
     
     
         45 - 74 . (canceled)