The polyspecific immunoglobulin response to HSV-1 viral proteins: Determination of immunogenic proteins and relative antibody titres to individual polypeptides by immunoblotting

R. R. McKendall, M. Pettit, W. Woo

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

We developed an immunoblotting procedure to characterise the polyspecific immunoglobulin response to the proteins of herpes simplex virus (HSV)-1. We found that 8-20% gradient polyacrylamide gels provided no advantage over fixed 8.5% gels for preparing Western blots for use in immunoblotting. The amount of protein loaded on the gels markedly influenced which proteins were detected by immune serum. The presence of Triton-X-100 0.5% in washes and buffers improved band clarity on immunoblots. In optimal conditions, immune mouse serum reacted with up to 33 HSV-1 lysate proteins. Six bands or regions appeared to be of major immunogenic reactivity, including the (122-130) x 103-mol. wt region, a 75 x 103-mol. wt protein, the gD region of approximately (56-64) x 103-mol. wt and two non-glycosylated bands at mol. wts(103) 42 and 44. Minor proteins, more weakly reactive, were detected at 27 other areas. The relative antibody titres in immune mouse serum to the different major regions showed antibody titre to gD>gB/gC>(42/44) x 103>>P75>>VP154. Most human sera reacted with all of the major and many of the minor immunogenic proteins but individual sera varied markedly in the proteins recognised. We conclude that immunoblotting is valuable for evaluating immunoglobulin responses to major and minor immunogenic proteins of HSV-1.

Original languageEnglish (US)
Pages (from-to)49-57
Number of pages9
JournalJournal of Medical Microbiology
Volume25
Issue number1
StatePublished - 1988

Fingerprint

Human Herpesvirus 1
Viral Proteins
Immunoblotting
Immunoglobulins
Peptides
Antibodies
Proteins
Immune Sera
Gels
Octoxynol
Blood Proteins
Buffers
Western Blotting
Serum

ASJC Scopus subject areas

  • Microbiology
  • Microbiology (medical)

Cite this

@article{645dc5322dd44c0e9977c9fda6cf321c,
title = "The polyspecific immunoglobulin response to HSV-1 viral proteins: Determination of immunogenic proteins and relative antibody titres to individual polypeptides by immunoblotting",
abstract = "We developed an immunoblotting procedure to characterise the polyspecific immunoglobulin response to the proteins of herpes simplex virus (HSV)-1. We found that 8-20{\%} gradient polyacrylamide gels provided no advantage over fixed 8.5{\%} gels for preparing Western blots for use in immunoblotting. The amount of protein loaded on the gels markedly influenced which proteins were detected by immune serum. The presence of Triton-X-100 0.5{\%} in washes and buffers improved band clarity on immunoblots. In optimal conditions, immune mouse serum reacted with up to 33 HSV-1 lysate proteins. Six bands or regions appeared to be of major immunogenic reactivity, including the (122-130) x 103-mol. wt region, a 75 x 103-mol. wt protein, the gD region of approximately (56-64) x 103-mol. wt and two non-glycosylated bands at mol. wts(103) 42 and 44. Minor proteins, more weakly reactive, were detected at 27 other areas. The relative antibody titres in immune mouse serum to the different major regions showed antibody titre to gD>gB/gC>(42/44) x 103>>P75>>VP154. Most human sera reacted with all of the major and many of the minor immunogenic proteins but individual sera varied markedly in the proteins recognised. We conclude that immunoblotting is valuable for evaluating immunoglobulin responses to major and minor immunogenic proteins of HSV-1.",
author = "McKendall, {R. R.} and M. Pettit and W. Woo",
year = "1988",
language = "English (US)",
volume = "25",
pages = "49--57",
journal = "Journal of Medical Microbiology",
issn = "0022-2615",
publisher = "Society for General Microbiology",
number = "1",

}

TY - JOUR

T1 - The polyspecific immunoglobulin response to HSV-1 viral proteins

T2 - Determination of immunogenic proteins and relative antibody titres to individual polypeptides by immunoblotting

AU - McKendall, R. R.

AU - Pettit, M.

AU - Woo, W.

PY - 1988

Y1 - 1988

N2 - We developed an immunoblotting procedure to characterise the polyspecific immunoglobulin response to the proteins of herpes simplex virus (HSV)-1. We found that 8-20% gradient polyacrylamide gels provided no advantage over fixed 8.5% gels for preparing Western blots for use in immunoblotting. The amount of protein loaded on the gels markedly influenced which proteins were detected by immune serum. The presence of Triton-X-100 0.5% in washes and buffers improved band clarity on immunoblots. In optimal conditions, immune mouse serum reacted with up to 33 HSV-1 lysate proteins. Six bands or regions appeared to be of major immunogenic reactivity, including the (122-130) x 103-mol. wt region, a 75 x 103-mol. wt protein, the gD region of approximately (56-64) x 103-mol. wt and two non-glycosylated bands at mol. wts(103) 42 and 44. Minor proteins, more weakly reactive, were detected at 27 other areas. The relative antibody titres in immune mouse serum to the different major regions showed antibody titre to gD>gB/gC>(42/44) x 103>>P75>>VP154. Most human sera reacted with all of the major and many of the minor immunogenic proteins but individual sera varied markedly in the proteins recognised. We conclude that immunoblotting is valuable for evaluating immunoglobulin responses to major and minor immunogenic proteins of HSV-1.

AB - We developed an immunoblotting procedure to characterise the polyspecific immunoglobulin response to the proteins of herpes simplex virus (HSV)-1. We found that 8-20% gradient polyacrylamide gels provided no advantage over fixed 8.5% gels for preparing Western blots for use in immunoblotting. The amount of protein loaded on the gels markedly influenced which proteins were detected by immune serum. The presence of Triton-X-100 0.5% in washes and buffers improved band clarity on immunoblots. In optimal conditions, immune mouse serum reacted with up to 33 HSV-1 lysate proteins. Six bands or regions appeared to be of major immunogenic reactivity, including the (122-130) x 103-mol. wt region, a 75 x 103-mol. wt protein, the gD region of approximately (56-64) x 103-mol. wt and two non-glycosylated bands at mol. wts(103) 42 and 44. Minor proteins, more weakly reactive, were detected at 27 other areas. The relative antibody titres in immune mouse serum to the different major regions showed antibody titre to gD>gB/gC>(42/44) x 103>>P75>>VP154. Most human sera reacted with all of the major and many of the minor immunogenic proteins but individual sera varied markedly in the proteins recognised. We conclude that immunoblotting is valuable for evaluating immunoglobulin responses to major and minor immunogenic proteins of HSV-1.

UR - http://www.scopus.com/inward/record.url?scp=0023931236&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023931236&partnerID=8YFLogxK

M3 - Article

C2 - 2826789

AN - SCOPUS:0023931236

VL - 25

SP - 49

EP - 57

JO - Journal of Medical Microbiology

JF - Journal of Medical Microbiology

SN - 0022-2615

IS - 1

ER -