Preparation and development of equine hyperimmune globulin F(ab')2 against severe acute respiratory syndrome coronavirus

Acta Pharmacol Sin. 2005 Dec;26(12):1479-84. doi: 10.1111/j.1745-7254.2005.00210.x.

Abstract

Aim: The resurgence of severe acute respiratory syndrome (SARS) is still a threat because the causative agent remaining in animal reservoirs is not fully understood, and sporadic cases continue to be reported. Developing high titers of anti-SARS hyperimmune globulin to provide an alternative pathway for emergent future prevention and treatment of SARS.

Methods: SARS coronavirus (CoV)F69 (AY313906) and Z2-Y3 (AY394989) were isolated and identified from 2 different Cantonese onset SARS patients. Immunogen was prepared from SARS-CoV F69 strain. Six health horses were immunized 4 times and serum was collected periodically to measure the profile of specific IgG and neutralizing antibodies using indirect enzyme-linked immunosorbent assay and a microneutralization test. Sera were collected in large amounts at the peak, where IgG was precipitated using ammonium sulphate and subsequently digested with pepsin. The product was then purified using anion-exchange chromatography to obtain F(ab')2 fragments.

Results: The specific IgG and neutralizing antibody titers peaked at approximately week 7 after the first immunization, with a maximum value of 1:14210. The sera collected at the peak were then purified. Fragment of approximately 15 g F(ab')2 was obtained from 1litre antiserum and the purity was above 90% with the titer of 1:5120, which could neutralize the other strain (SARS-CoV Z2-Y3) as well.

Conclusion: This research provides a viable strategy for the prevention and treatment of SARS coronavirus infection with equine hyperimmune globulin, with the purpose of combating any resurgence of SARS.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Viral / chemistry
  • Antibodies, Viral / isolation & purification
  • Enzyme-Linked Immunosorbent Assay
  • Horses
  • Humans
  • Immune Sera / immunology*
  • Immunization
  • Immunoglobulin Fab Fragments / chemistry
  • Immunoglobulin Fab Fragments / isolation & purification*
  • Immunoglobulin G / chemistry
  • Immunoglobulin G / isolation & purification*
  • Neutralization Tests
  • Severe Acute Respiratory Syndrome / virology
  • Severe acute respiratory syndrome-related coronavirus / immunology*
  • Severe acute respiratory syndrome-related coronavirus / isolation & purification

Substances

  • Antibodies, Viral
  • Immune Sera
  • Immunoglobulin Fab Fragments
  • Immunoglobulin G