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检测p2 4抗原的诊断试剂在AIDS研究和防治方面具有重要意义。为研制国产HIVp2 4抗原诊断试剂 ,根据国内外文献 ,合成了HIVp2 4抗原的六个肽段 :G -A 12 ,D -R 16 ,P -S 18,A -G 2 3(HIV - 2ROD) ,P -S 18,N -I 15以及D -C 2 2。六个肽段中 ,N -I 15肽和D -C 2 2肽用于制备McAb ,其余 4个用于制备山羊抗血清。除D -R 16肽的滴度较低 (1∶2 0 0 0 )外 ,其余三个肽的抗血清滴度都在 1∶10 0 0 0以上。McAb中 ,N -I 15肽的反应比D -C 2 2肽要强 ,但两者的腹水滴度相同 ,均为 1∶10 0 0。单抗铺板检测病毒 p2 4抗原的非特异性反应比较强。用山羊抗血清铺板 ,1∶80 0 0的稀释度检测效果最好。用我们研制的抗体检测不出裂解HIV - 1ⅢB中 p2 4 ,却能够测出Abbott公司p2 4抗原诊断试剂盒的阳性对照 (HIVAG - 1)。用北海道大学免疫科学研究所研制的单抗和人阳性血清做的交叉对照实验表明 ,可能是由于合成肽免疫产生的抗体与裂解病毒p2 4的亲和力较差引起
Diagnostic reagents for the detection of p24 antigen are of great importance in AIDS research and prevention. To develop domestic HIVp2 4 antigen diagnostic reagents, six peptides of HIVp2 4 antigen were synthesized according to domestic and foreign literatures: G-A12, D-R16, P-S18 and A-G22 , P-S 18, N-I 15, and D-C 2 2. Of the six peptides, N-I 15 peptide and D-C 2 2 peptide were used to prepare McAb and the remaining 4 were used to prepare goat antisera. The antiserum titers of the remaining three peptides were above 1: 10000, except for the low titer of the D-R16 peptide (1: 200). In the McAb, the N-I 15 peptide reacted more strongly than the D-C 2 2 peptide, but the ascites titers of both were the same, both of which were 1:10 0 0. Monoclonal antibody detection of p24 antigen-specific non-specific anti-virus response is relatively strong. With goat antiserum plating, dilution of 1: 80000 is the best. Using the antibodies we developed, no detectable cleavage of p2 4 in HIV - 1 Ⅲ B was able to detect a positive control (HIVAG - 1) from Abbott ’s p2 4 antigen diagnostic kit. Cross-control experiments with monoclonal and human positive sera developed by the Institute of Immunology, Hokkaido University showed that the antibodies, possibly due to the immunization with synthetic peptides, are less likely to be associated with the lytic virus p24