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- Title
模拟表位结合夹心化学发光法检测抗 A 和抗 B 效价.
- Authors
周水梅; 沈长新; 宋晶晶; 王娇; 林晨瑶; 黄爽; 贺学宇
- Abstract
Objective: The aim of this study was to acquire the mimic peptides of A/B antigen and combined it with sandwich chemiluminescence immunoassay (CLIA)to develop more effective way to measure the titer of A/B antibody. Methods: Three-round of biopanning was performed on the phage 12 peptide library with NaM87-1F6 and HEB-29 as targets,respectively. Positive plaques determined by enzyme-linked immunosorbent assay (ELISA) were then sequenced, and the mimetic peptides were synthesized according to their amino acid sequences. Competitive ELISA and agglutination inhibition were carried out to confirm the interaction between anti-A/B and the synthesized peptides. The mimetic peptides was then combined with CLIA to detect A and B antibody titers. Standard curves were prepared and the results obtained by the method based on CLIA and the micro-column agglutination technique (CAT) method were compared and the correlation between them was analyzed. Results: After three rounds of panning and related identification, 13 and 19 phage clones that specifically interacted with anti-A and anti-B antibodies were obtained, of which 9(MTRIQLRMMRLH) and 12 (PQMSRHRMRMLP) showed the same amino acid sequences, respectively. The synthesized peptides were named MTR and PQM, respectively. The results of competitive ELISA indicated that MTR can specifically antagonize the binding of anti- A and the replacement polypeptide of the A antigen; PQM can specifically antagonize the immunological binding of anti-B and the replacement polypeptide of the B antigen. Agglutination inhibition experiments showed that MTR/PQM could specifically inhibit the interaction between group A/B red blood cells and anti-A/B. The synthesized peptides combined with CLIA method proposed here had a broad linear range (2-2048) and a good linear correlation coefficient (for A antibody R2=0.9566, for B antibody R2=0.9762). The results obtained by the method based on CLIA were highly correlated with the results obtained by the CAT method (P<0.001). Conclusions: MTR and PQM have good A or B antigenicity and have the potential to be the artificial replacements of them. The method based on the combination of mimic peptides and the CLIA can quantitatively detect the titer of A and B antibodies, and the results are accurate and reliable.
- Subjects
ADENOSYLMETHIONINE; AMINO acid sequence; CHEMILUMINESCENCE immunoassay; ENZYME-linked immunosorbent assay; ERYTHROCYTES; TITERS
- Publication
Progress in Modern Biomedicine, 2019, Vol 19, Issue 15, p2822
- ISSN
1673-6273
- Publication type
Article
- DOI
10.13241/j.cnki.pmb.2019.15.005