Detection of IgM and IgA HIV antibodies after removal of IgG with recombinant protein G

J Immunol Methods. 1990 Feb 9;126(2):199-204. doi: 10.1016/0022-1759(90)90151-k.

Abstract

Indirect assays for IgM and IgA antibodies often lack sensitivity and specificity due to interference from IgG antibodies. To overcome this problem we have developed a simple procedure using recombinant protein G coupled to agarose beads to remove the interfering IgG. A series of HIV seroconversion panels was tested by IgM and IgA immunoblot after protein G treatment in order to evaluate IgG removal and to study appearance of IgM and IgA antibodies in early HIV infection. Protein G treatment removed 99.9% of the IgG and reduced IgG anti-HIV titers of over 1/100,000 to undetectable levels. Both IgM and IgA HIV antibodies were detected as early in seroconversion as were IgG HIV antibodies. IgA HIV antibodies persisted for a longer period of time, reacted with more HIV proteins, and showed more intense staining than IgM HIV antibodies.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Bacterial Proteins / metabolism*
  • Enzyme-Linked Immunosorbent Assay
  • Evaluation Studies as Topic
  • HIV Antibodies / analysis*
  • HIV Seropositivity / immunology*
  • Humans
  • Immunoblotting
  • Immunoglobulin A / analysis
  • Immunoglobulin A / immunology*
  • Immunoglobulin G / metabolism*
  • Immunoglobulin M / analysis
  • Immunoglobulin M / immunology*
  • Immunologic Techniques
  • Recombinant Proteins
  • Sepharose

Substances

  • Bacterial Proteins
  • HIV Antibodies
  • IgG Fc-binding protein, Streptococcus
  • Immunoglobulin A
  • Immunoglobulin G
  • Immunoglobulin M
  • Recombinant Proteins
  • Sepharose