Skip to main content

Advertisement

SpringerLink
Log in

Preparation of Immunoglobulin Y (IgY) Against Lipopolysaccharide using Gel Chromatography from the Yolks of Eggs Laid by Immunized Hens

  • Published:
The Protein Journal Aims and scope Submit manuscript

Abstract

The objective is to prevent and treat injuries caused by lipopolysaccharide (LPS) from gram negative bacteria in animals and humans, we produced antibodies against LPS from egg yolk. LPS from E. coli (O111:B4) mixed with Freund’s Adjuvant was used as the immunogen to immunize Roman hens. Immunized eggs were collected, and immunoglobulin Y (IgY) was purified using a water solution, salt precipitation and gel chromatography. The molecular weight and purity were determined by SDS–PAGE, the antibody titer by noncompetitive enzyme-linked immunosorbent assay (ELISA) and antibody activity against LPS by the mortality of mice intraperitoneally injected with LPS or LPS-IgY solutions. IgY against LPS showed two protein bands at 68 and 26 kDa on the gel; the antibody titer was almost 1:25,600. After incubation with LPS, IgY decreased the mortality of mice challenged with LPS. This study provided an efficient way to produce high-titer egg yolk antibodies, which could attenuate lethal effects of LPS, by immunizing hens. Furthermore, the LPS antibody was purified well using a water solution, salting-out and gel chromatography.

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

Similar content being viewed by others

Abbreviations

IgY:

Immunoglobulin Y; egg yolk antibody

LPS:

Lipopolysaccharide

E. coli :

Escherichia coli

HCl:

Hydrogen chloride

Na2SO4 :

Sodium sulfate

OD:

Optical density

PBS:

Phosphate-buffered saline

References

  1. Akita EM, Nakai S (1993) J Immunol Methods 160:207–214

    Article  CAS  Google Scholar 

  2. Cross AS, Opal SM, Palardy JE, Drabick JJ, Warren HS, Huber C, Cook P, Bhattacharjee AK (2003) Vaccine 21:4576–4587

    Article  CAS  Google Scholar 

  3. Amaral JA, De Franco MT, Zapata-Quintanilla L, Carbonare SB (2008) Vet Res Commun 32:281–290

    Article  Google Scholar 

  4. Bhattacharjee AK, Opal SM, Palardy JE, Drabick JJ, Collins H, Taylor R, Cotton A, Cross AS (1994) J Infect Dis 70:622–629

    Google Scholar 

  5. Chalghoumi R, Thewis A, Beckers Y, Marcq C, Portetelle D, Schneider YJ (2009) Foodborne Pathog Dis 6:593–604

    Article  CAS  Google Scholar 

  6. Raetz C, Whitfield C (2002) Annu Rev Biochem 71:635–700

    Article  CAS  Google Scholar 

  7. de Almeida CMC, Da Silva CL, Couto HP, de Escocard RCM, da Rocha DG, de Paula Sentinelli L, Kipnis TL, da Silva WD (2008) Toxicon 52:293–301

    Article  Google Scholar 

  8. Derkx B, Witter J, Mccloskey R (1999) Clin Infect Dis 28:770–777

    Article  CAS  Google Scholar 

  9. Di Padova FE, Brade H, Barclay GR, Poxton IR, Liehl E, Schuetze E, Kocher HP, Ramsay G, Schreier MH, McClelland DB (1993) Infect Immun 61:3863–3872

    CAS  Google Scholar 

  10. Fomsgaard A, Baek L, Fomsgaard JS, Engquist A (1989) Scand J Infect Dis 21:697–708

    Article  CAS  Google Scholar 

  11. Yi-bin G, Hong-wei C, Liping C, Yong-ling L, Jiang Z, Guang-xia X (2007) Chin Jl Burns 23:104–107

    Google Scholar 

  12. Harkin DW, Barros D’Sa AA, Yassin MM, Hoper M, Halliday MI, Parks TG, Campbell FC (2001) J Vas Surg 33:840–846

    Article  CAS  Google Scholar 

  13. Amaral JA, De Franco MT, Zapata-Quintanilla L, Carbonare SB (2008) Vet Res Commun 32:281–290

    Article  Google Scholar 

  14. Krief A, Letesson JJ, Billen D (2002) Tetrahedron Lett 43:1843–1846

    Article  CAS  Google Scholar 

  15. Lachman E, Pitsoe SB, Gaffin SL (1984) Lancet 1:981–983

    Article  CAS  Google Scholar 

  16. Le Roy D, Di Padova F, Tees R, Lengacher S, Landmann R, Glauser MP, Calandra T, Heumann D (1999) J Immunol 162:7454–7460

    Google Scholar 

  17. Mayumi T, Takezawa J, Takahashi H, Kuwayama N, Fukuoka T, Shimizu K, Yamada K, Kondo S, Aono K (1999) Shock 11:82–86

    Article  CAS  Google Scholar 

  18. MT Madigan, JM Martinko, J Parker (2003) In: Brock biology of microorganisms. Pearson-Prentice Hall, Madrid, 1012 pp. ISBN 84-205-3679-2

  19. Nomura S, Suzuki H, Masaoka T, Kurabayashi K, Ishii H, Kitajima M, Nomoto K, Hibi T (2005) Helicobacter 10:43–52

    Article  CAS  Google Scholar 

  20. van Langevelde P, Kwappenberg KMC, Groeneveld PHP, Mattie H, van Dissel JT (1998) Antimicrob Agents Chemother 42:739–743

    Google Scholar 

  21. Shi z, Zhang Y, Ma S (2009) Immunol J 25:337–340

    CAS  Google Scholar 

  22. Steven AM, James PT (2001) Biochemisty 40:5777–5785

    Article  Google Scholar 

  23. Suzuki H, Nomura S, Masaoka T, Goshima H, Kamata N, Kodama Y, Ishii H, Kitajima M, Nomoto K, Hibi T (2004) Aliment Pharmacol Ther 20(Suppl 1):185–192

    Article  CAS  Google Scholar 

  24. Tsuji M, Matsuda H, Miwa H, Miyazaki S (2003) J Antimicrob Chemother 51:353–359

    Article  CAS  Google Scholar 

  25. Lu Y, Liu J, Jin L, Li X, Zhen YH, Xue H, You J, Xu Y (2008) Fish Shellfish Immunol 25:604–610

    Article  CAS  Google Scholar 

  26. Zhen YH, Jin LJ, Li XY, Guo J, Li Z, Zhang BJ, Fang R, Xu YP (2009) Vet Microbiol 133:317–322

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors thank the staff of the State Key Laboratory of Trauma, Burns and Combined Injury, Southwest Hospital, Third Military Medical University, for their assistance in caring for the animals, collecting eggs and providing technical services used in this study.

Author information

Authors and Affiliations

  1. Institute of Burn Research, Southwest Hospital, State Key Laboratory of Trauma, Burns and Combined Injury, The Third Military Medical University, Gao Tanyan Main Street 30th, 400038, Shapingba District, Chongqing, People’s Republic of China

    Siyuan Ma & Yaping Zhang

Authors
  1. Siyuan Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yaping Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yaping Zhang.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ma, S., Zhang, Y. Preparation of Immunoglobulin Y (IgY) Against Lipopolysaccharide using Gel Chromatography from the Yolks of Eggs Laid by Immunized Hens. Protein J 29, 475–480 (2010). https://doi.org/10.1007/s10930-010-9275-z

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10930-010-9275-z

Keywords

Search

Navigation