Skip to main content

Advertisement

SpringerLink
Log in

Secretory expression of a novel human spermatozoa antigen in E. coli and its application to a protein chip

  • Original Research Paper
  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

Objective

To produce a recombinant spermatozoa antigen peptide using the E. coli: PhoA system on a protein chip for screening anti-sperm antibodies (ASA).

Results

The purity of the recombinant spermatozoa antigen exceeded 95% after two-step purification, as assessed using SDS-PAGE and HPLC. The diagnostic performance of a protein chip coated with the recombinant antigen peptide was evaluated by examining ASA in 51 infertile patients in comparison with a commercial ELISA kit. The area under the receiver operating characteristic curve (AUC) was 0.944, which indicated that the protein chip coated with recombinant spermatozoa antigen peptide was consistent with ELISA for ASA detection.

Conclusion

A recombinant spermatozoa antigen was expressed in the E. coli PhoA secretory expression system and its potential application for clinical ASA detection was validated.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Brincat D, Catania S, Wismayer PS, Calleja-Agius J (2015) Male factors in ART outcome prediction. Gynecol Endocrinol 31:169–175

    Article  PubMed  Google Scholar 

  • Campion SR, Elsasser E, Chung R (1997) Amino-terminal charge affects the periplasmic accumulation of recombinant heregulin/EGF hybrids exported using the Escherichia coli alkaline phosphatase signal sequence. Protein Expr Purif 10:331–339

    Article  CAS  PubMed  Google Scholar 

  • Choi JH, Lee SY (2004) Secretory and extracellular production of recombinant proteins using Escherichia coli. Appl Microbiol Biotechnol 64:625–635

    Article  CAS  PubMed  Google Scholar 

  • Cui D, Han G, Shang Y, Liu C, Xia L, Li L, Yi S (2015) Antisperm antibodies in infertile men and their effect on semen parameters: a systematic review and meta-analysis. Clin Chim Acta 444:29–36

    Article  CAS  PubMed  Google Scholar 

  • Domagała A, Kamieniczna M, Kurpisz M (2000) Sperm antigens recognized by antisperm antibodies present in sera of infertile adults and prepubertal boys with testicular failure. Int J Androl 23:150–155

    Article  PubMed  Google Scholar 

  • Eom GT, Lee SH, Oh YH, Choi JE, Park SJ, Song JK (2014) Efficient extracellular production of type I secretion pathway-dependent Pseudomonas fluorescens lipase in recombinant Escherichia coli by heterologous ABC protein exporters. Biotechnol Lett 36:2037–2042

    Article  CAS  PubMed  Google Scholar 

  • Fernandes LG et al (2017) Immune response and protective profile elicited by a multi-epitope chimeric protein derived from Leptospira interrogans. Int J Infect Dis 57:61–69

    Article  CAS  PubMed  Google Scholar 

  • Lu JC, Huang YF, Lu NQ (2008) Antisperm immunity and infertility. Exp Rev Clin Immunol 4:113–126

    Article  CAS  Google Scholar 

  • Qi L et al (2017) Development of a highly specific HER2 monoclonal antibody for immunohistochemistry using protein microarray chips. Biochem Biophys Res Commun 484:248–254

    Article  CAS  PubMed  Google Scholar 

  • Selvarajah S et al (2014) Development and validation of protein microarray technology for simultaneous inflammatory mediator detection in human sera. Mediat Inflamm 2014:820304

    Article  Google Scholar 

  • Sikka SC, Hellstrom WJ (2016) Current updates on laboratory techniques for the diagnosis of male reproductive failure. Asian J Androl 18:392–401

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Su Q et al (2016) Epitope-based recombinant diagnostic antigen to distinguish natural infection from vaccination with hepatitis A virus vaccines. J Virol Methods 233:41–45

    Article  PubMed  Google Scholar 

  • Vazquez-Levin MH, Marin-Briggiler CI, Veaute C (2014) Antisperm antibodies: invaluable tools toward the identification of sperm proteins involved in fertilization. Am J Reprod Immunol 72:206–218

    Article  CAS  PubMed  Google Scholar 

  • Xu R, Du P, Fan JJ, Zhang Q, Li TP, Gan RB (2002) High-level expression and secretion of recombinant mouse endostatin by Escherichia coli. Protein Expr Purif 24:453–459

    Article  CAS  PubMed  Google Scholar 

  • Xu QY et al (2016) Development of a novel protein chip for the detection of bluetongue virus in China. J Virol Methods 234:28–33

    Article  CAS  PubMed  Google Scholar 

Download references

Funding

This study was supported by Science and Technology Program of Guangzhou (Grant No. 201604020104), Science and Technology Program of Guangdong (Grant No. 2015A030401040) and the Key Program for health care collaborative innovation of Guangzhou (Grant No. 201500000004-4).

Author information

Authors and Affiliations

  1. Institute of Antibody Engineering, School of Laboratory Medical and Biotechnology, Southern Medical University, Guangzhou, People’s Republic of China

    Ming-zhu Wang, Xiang-Sheng Cai, Jing-jing Li & Ying-song Wu

  2. Reproductive center of Obstetrics and Gynecology, Southern Medical University, Guangzhou, 510515, People’s Republic of China

    Zhuo-lin Qiu

  3. Guangzhou Kangsui Biotechnology Inc., Guangzhou, 510663, People’s Republic of China

    Miao-qin She & Yuan-feng Xu

Authors
  1. Ming-zhu Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zhuo-lin Qiu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xiang-Sheng Cai
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jing-jing Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Miao-qin She
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yuan-feng Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Ying-song Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ying-song Wu.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wang, Mz., Qiu, Zl., Cai, XS. et al. Secretory expression of a novel human spermatozoa antigen in E. coli and its application to a protein chip. Biotechnol Lett 39, 1529–1535 (2017). https://doi.org/10.1007/s10529-017-2377-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10529-017-2377-3

Keywords

Search

Navigation