[Preparation of high-affinity monclonal antibody against hemoglobin delta globin chain and beta globin chain].
OBJECTIVE
To prepare monclonal antibody (mAb) against both hemoglobin A2 (HbA2) and hemglobin A (HbA), this antiboy can bind to delta globin chain and beta globin chain, but not to gamma globin chain.


METHODS
Mice was immunized with recombinant Hb delta fusion protein, and hybridoma cells were generated by cell fusion techniques followed by screening with natural HbA2 and HbA separated by ion exchange chromatography. The purified monclonal antibody was identified by indirect ELISA, Western blot based on native PAGE and SDS-PAGE, surface plasmon resonance (SPR), flow cytometry and immunohistochemistry.


RESULTS
The monoclonal antibody against both HbA2 and HbA was obtained and designated as 2C9 that shows no binding to fetal hemoglobin (HbF), alpha globin chain and recombinant zetaglobin chain.


CONCLUSIONS
The mAb 2C9 was defined as specificity to hemoglobin delta globin chain and beta globin chain, which suggests that mAb 2C9 recognizes a common epitope on bothdeltaglobin chain and beta globin chain. This antibody would be expected to be an effective tool in research and clinical practice in hemoglobinopathies.
Zhu
Ping
P
Department of Immunology, School of Basic Medical Science, Southern Medical University, Guangzhou 510515, China. zhuping@finmu.com
Xiao
Hai-yan
HY
Chen
Yi-guo
YG
Xu
Mei
M
Fu
Ning
N
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology*; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006443:Hemoglobin A2 / Q000276:immunology; D006454:Hemoglobins / Q000276:immunology; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D055542:alpha-Globins / Q000276:immunology; D055544:beta-Globins / Q000276:immunology; D055546:delta-Globins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D006454:Hemoglobins; D055542:alpha-Globins; D055544:beta-Globins; D055546:delta-Globins; D006443:Hemoglobin A2