Generation and evaluation of antibodies against human MGF E-peptide by reverse phase protein microarray and reverse competitive ELISA.
BACKGROUND
Since 2005, as one of prohibited substances on the Prohibited List of the World Anti-Doping Agency (WADA), the occurence of mechano growth factor (MGF) abuse in sport has likely increased. However, there is still no WADA-validated and -approved method for its detection.


RESULTS
Four polyclonal antibodies (Ab-K01, Ab-B01, Ab-B02 and Ab-K02) against MGF C-terminal peptides were generated, purified and evaluated by western blot, ELISA and reverse-phase protein microarray, respectively. It was found that all the antibodies could identify their corresponding antigen in mouse serum by reverse-phase protein microarray, in particular, Ab-K01 showed the highest affinity among them and might be a potential tool for the detection of antibody affinity. Furthermore, Ab-B01 and Ab-K01 were successfully used for the determination of MGF-40 by reverse competitive ELISA.


CONCLUSIONS
The quantitative measurement of MGF-40 has laid the foundation for doping detection of MGF and further biological research on MGF.
Department of Biotechnology, School of Chemistry & Biological Engineering, University of Science & Technology Beijing, 30 Xueyuan Rd, Beijing 100083, China.
Hongwu
H
Du
Chuanbao
C
Li
Haiming
H
Jin
Guangyu
G
Chen
Yiping
Y
Xun
D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000276:immunology*; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions / Q000276:immunology; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000032:analysis* / Q000235:genetics / Q000276:immunology*; D051379:Mice; D040081:Protein Array Analysis; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology
D000906:Antibodies; D011994:Recombinant Proteins; C523997:mechano-growth factor E, human; D007334:Insulin-Like Growth Factor I