Generation and evaluation of antibodies against human MGF E-peptide by reverse phase protein microarray and reverse competitive ELISA. (September 2013)

• Record Type: Journal Article
• Authors: Du, Hongwu; Li, Chuanbao; Jin, Haiming; Chen, Guangyu; Xun, Yiping
• Is Part Of: Bioanalysis. Volume 5:Number 18(2013)
• Page Start: 2269
• Page End: 2275
• Publication Date: 2013-09
• Subjects: Drugs -- Analysis -- Periodicals; 615.19005
• DOI: 10.4155/bio.13.195
• Languages: English
• ISSNs: 1757-6180

Abstract

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.Conclusion: The quantitative measurement of MGF-40 has laid the foundation for doping detection of MGF and further biological research on MGF.