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KMID : 0578319960060020169
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Molecules and Cells
1996 Volume.6 No. 2 p.169 ~ p.175
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Use of Multiple Antigenic Peptides as Coating Antigens in Detection of Antibody
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Shin, Song Yub
Lee, Myung Kyu/Kim, Sun Young/Hahm, Kyung-Soo
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Abstract
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The protein conjugates of two peptides (P1 and P2) with the sequences 101-116 and 253-274 of the human immunodeficiency virus type 1 (HIV1) gpl20 protein, respectively, showed high reactivity against an anti-gpl20 goat antibody, whereas the peptides only displayed low reactivity when used as coating antigens in indirect enzyme linked immuno sorbent assay (ELISA). However, P1 and P2 effectively inhibited the binding of the anti-HIV1 gpl20 goat antibody with their peptide-ovalbumin conjugates in competitive ELISA, suggesting that the low reactivity of P1 and P2 in indirect ELISA resulted from their low coating efficiency on the microtiter plates. To increase coating efficiency of synthetic peptides without conjugation, four multiple antigenic peptides (MAP) for each peptide were synthesized. Their antigenicity was determined against the anti-HIV1 gpl20 goat antibody and anti-peptide antisera by indirect and competitive ELISA. The MAPs for both P1 and P2 showed almost similar reactivity with the peptide-bovine serum albumin conjugates in indirect ELISA. MAPs with four peptide branches were more effective than MAPs with two peptide branches. The high efficiency of the MAPs seems to be related to the cumulative effects of their high coating efficiency and high affinity conformation on the microtiter plate to the antibodies. These results suggest that multi-branched MAPs are useful as coating peptide antigens in the development of analytical systems for disease diagnosis.
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KEYWORD
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