Characterization of one monoclonal antibody against feline immunodeficiency virus p24 and its application to antigen capture ELISA.
Several monoclonal antibodies (mAbs) were produced against feline immunodeficiency virus (FIV) p24 capsid antigen. One of these, F2710, reacted strongly, not only with viral p24 and recombinant p24, but also with p50 Gag precursor protein in Western blot. Epitope mapping analysis revealed that mAb F2710 recognizes a heptapeptide, SFIDRLF, in the FIV p24 amino acid sequence. As this portion of FIV p24 is highly conserved among various FIV strains, the mAb seems to be a useful tool for detecting FIV p24 antigen in various samples. By means of this mAb and rabbit anti-p24 polyclonal antibody, an antigen capture ELISA was developed. The ELISA detected viral p24 antigen with good linearity. The lower detection limit of this assay is 40 pg/ml of recombinant p24 antigen, which is at least as sensitive as the reverse transcriptase assay in detecting FIV virion. Thus, this system is valuable for monitoring FIV replication in vitro.
Kashiwase
H
H
Biological Research Laboratories, Sankyo Co., Ltd., Tokyo, Japan. hiroto@shina.sankyo.co.jp
Ishimura
M
M
Ishikawa
Y
Y
Nishigaki
T
T
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry*; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D002415:Cats; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D015683:Gene Products, gag / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D007106:Immune Sera / Q000096:biosynthesis / Q000737:chemistry; D016089:Immunodeficiency Virus, Feline / Q000276:immunology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009101:Multiple Myeloma; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D015683:Gene Products, gag; D007106:Immune Sera; D011994:Recombinant Proteins; C074662:p24 protein, Feline immunodeficiency virus