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Detection of Antibodies to HCV E1E2 by Lectin-Capture ELISA

  • Marian MajorEmail author
  • Mansun Law
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1911)

Abstract

Enzyme-linked immunosorbent assays (ELISAs) enable rapid detection and quantitation of antibodies in samples. Such assays can be highly sensitive and can be performed in most laboratories with basic equipment. Although detecting binding antibodies to the surface proteins of most pathogens by ELISA is not always indicative of antibody function, i.e., neutralizing activity of antibodies, the results can be used as a first step toward more in-depth analysis of antibody responses. Here we describe a method that can be used to standardize ELISAs for the detection of HCV envelope antibodies across laboratories and provide adaptations of the method to further characterize antibody responses in serum samples.

Key words

ELISA Lectin Glycoproteins Antibody titer Affinity Avidity 

Notes

Acknowledgments

Financial support was provided by Food and Drug Administration intramural funds [Program Number Z01 BK 04010-11 LHV to M.M.] and by the National Institutes of Health [grant numbers AI079037, AI106005 and AI123861 to M.L.]. We thank Yusra Gimie and Kenna Nagy for comments and proofreading of the manuscript.

References

  1. 1.
    Logvinoff C, Major ME, Oldach D, Heyward S, Talal A, Balfe P et al (2004) Neutralizing antibody response during acute and chronic hepatitis C virus infection. Proc Natl Acad Sci U S A 101:10149–10154CrossRefGoogle Scholar
  2. 2.
    Pestka JM, Zeisel MB, Blaser E, Schurmann P, Bartosch B, Cosset FL et al (2007) Rapid induction of virus-neutralizing antibodies and viral clearance in a single-source outbreak of hepatitis C. Proc Natl Acad Sci U S A 104:6025–6030CrossRefGoogle Scholar
  3. 3.
    Dowd KA, Netski DM, Wang XH, Cox AL, Ray SC (2009) Selection pressure from neutralizing antibodies drives sequence evolution during acute infection with hepatitis C virus. Gastroenterology 136:2377–2386CrossRefGoogle Scholar
  4. 4.
    Osburn WO, Snider AE, Wells BL, Latanich R, Bailey JR, Thomas DL et al (2014) Clearance of hepatitis C infection is associated with the early appearance of broad neutralizing antibody responses. Hepatology 59:2140–2151CrossRefGoogle Scholar
  5. 5.
    Houghton M, Abrignani S (2005) Prospects for a vaccine against the hepatitis C virus. Nature 436:961–966CrossRefGoogle Scholar
  6. 6.
    Law M, Maruyama T, Lewis J, Giang E, Tarr AW, Stamataki Z et al (2008) Broadly neutralizing antibodies protect against hepatitis C virus quasispecies challenge. Nat Med 14:25–27CrossRefGoogle Scholar
  7. 7.
    Hsu M, Zhang J, Flint M, Logvinoff C, Cheng-Mayer C, Rice CM et al (2003) Hepatitis C virus glycoproteins mediate pH-dependent cell entry of pseudotyped retroviral particles. Proc Natl Acad Sci U S A 100:7271–7276CrossRefGoogle Scholar
  8. 8.
    Owsianka A, Tarr AW, Juttla VS, Lavillette D, Bartosch B, Cosset FL et al (2005) Monoclonal antibody AP33 defines a broadly neutralizing epitope on the hepatitis C virus E2 envelope glycoprotein. J Virol 79:11095–11104CrossRefGoogle Scholar
  9. 9.
    Keck Z, Wang W, Wang Y, Lau P, Carlsen TH, Prentoe J et al (2013) Cooperativity in virus neutralization by human monoclonal antibodies to two adjacent regions located at the amino terminus of hepatitis C virus E2 glycoprotein. J Virol 87:37–51CrossRefGoogle Scholar
  10. 10.
    Wong JA, Bhat R, Hockman D, Logan M, Chen C, Levin A et al (2014) Recombinant hepatitis C virus envelope glycoprotein vaccine elicits antibodies targeting multiple epitopes on the envelope glycoproteins associated with broad cross-neutralization. J Virol 88:14278–14288CrossRefGoogle Scholar
  11. 11.
    Helle F, Wychowski C, Vu-Dac N, Gustafson KR, Voisset C, Dubuisson J (2006) Cyanovirin-N inhibits hepatitis C virus entry by binding to envelope protein glycans. J Biol Chem 281:25177–25183CrossRefGoogle Scholar
  12. 12.
    Kachko A, Loesgen S, Shahzad-Ul-Hussan S, Tan W, Zubkova I, Takeda K et al (2013) Inhibition of hepatitis C virus by the cyanobacterial protein MVL: mechanistic differences between the high-mannose specific lectins MVL, CV-N, and GNA. Mol Pharm 10:4590–4602CrossRefGoogle Scholar
  13. 13.
    Pohlmann S, Zhang J, Baribaud F, Chen Z, Leslie GJ, Lin G et al (2003) Hepatitis C virus glycoproteins interact with DC-SIGN and DC-SIGNR. J Virol 77:4070–4080CrossRefGoogle Scholar
  14. 14.
    Takebe Y, Saucedo CJ, Lund G, Uenishi R, Hase S, Tsuchiura T et al (2013) Antiviral lectins from red and blue-green algae show potent in vitro and in vivo activity against hepatitis C virus. PLoS One 8:e64449CrossRefGoogle Scholar
  15. 15.
    Choo QL, Kuo G, Ralston R, Weiner A, Chien D, Van Nest G et al (1994) Vaccination of chimpanzees against infection by the hepatitis C virus. Proc Natl Acad Sci U S A 91:1294–1298CrossRefGoogle Scholar
  16. 16.
    Puig M, Major ME, Mihalik K, Yu MY, Feinstone SM (2004) Immunization of chimpanzees with an envelope protein-based vaccine enhances specific humoral and cellular immune responses that delay hepatitis C virus infection. Vaccine 22:991–1000CrossRefGoogle Scholar
  17. 17.
    Khan AG, Whidby J, Miller MT, Scarborough H, Zatorski AV, Cygan A et al (2014) Structure of the core ectodomain of the hepatitis C virus envelope glycoprotein 2. Nature 509:381–384CrossRefGoogle Scholar
  18. 18.
    Kong L, Giang E, Nieusma T, Kadam RU, Cogburn KE, Hua Y et al (2013) Hepatitis C virus E2 envelope glycoprotein core structure. Science 342:1090–1094CrossRefGoogle Scholar
  19. 19.
    Whidby J, Mateu G, Scarborough H, Demeler B, Grakoui A, Marcotrigiano J (2009) Blocking hepatitis C virus infection with recombinant form of envelope protein 2 ectodomain. J Virol 83:11078–11089CrossRefGoogle Scholar
  20. 20.
    Hacker DL, Balasubramanian S (2016) Recombinant protein production from stable mammalian cell lines and pools. Curr Opin Struct Biol 38:129–136CrossRefGoogle Scholar
  21. 21.
    Michalak JP, Dubuisson J, Rice CM, Ung S, Meunier JC, Choukhi A et al (1997) Characterization of truncated forms of hepatitis C virus glycoproteins. J Gen Virol 78:2299–2306CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Division of Viral Products, Center for Biologics Evaluation and ResearchFood and Drug AdministrationSilver SpringUSA
  2. 2.Department of Immunology and MicrobiologyThe Scripps Research InstituteLa JollaUSA

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