Potent malaria transmission-blocking antibody responses elicited by Plasmodium falciparum Pfs25 expressed in Escherichia coli after successful protein refolding

Rajesh Kumar, Evelina Angov, Nirbhay Kumar

Research output: Contribution to journalArticle

Abstract

Production of Pfs25, a Plasmodium falciparum transmission-blocking vaccine target antigen, in functional conformation with the potential to elicit effective immunogenicity still remains a major challenge. In the current study, codon-harmonized recombinant Pfs25 (CHrPfs25) was expressed in Escherichia coli, and purified protein after simple oxidative refolding steps retained reduction-sensitive conformational epitopes of transmission-blocking monoclonal antibodies. CHrPfs25 formulated in several adjuvants elicited strong immunogenicity in preclinical studies in mice. Antibodies elicited after immunization recognized native Pfs25 on the surface of live gametes of P. falciparum and demonstrated complete malaria transmission-blocking activity. The transmission-blocking efficacy was 100% even after a 1:128 dilution of sera from immunized mice in the complete Freund's adjuvant and Montanide ISA51 groups and after a 1:16 dilution of sera from mice in the alum group. The blocking was mediated by antibodies; purified IgG at concentrations as low as 31.25 μg/ml exhibited 100% transmission blocking in membrane feeding assays employing two different species of mosquitoes, Anopheles gambiae and Anopheles stephensi. This study provides the first evidence for successful expression of biologically functional rPfs25 in E. coli. The extremely potent malaria transmission-blocking activity of antibodies elicited by immunization with purified protein provides strong support for further evaluation of E. coli-derived CHrPfs25 as a malaria transmission-blocking vaccine in human clinical trials.

Original languageEnglish (US)
Pages (from-to)1453-1459
Number of pages7
JournalInfection and Immunity
Volume82
Issue number4
DOIs
StatePublished - 2014
Externally publishedYes

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Protein Refolding
Blocking Antibodies
Plasmodium falciparum
Codon
Malaria
Antibody Formation
Escherichia coli
Immunization
Vaccines
Anopheles gambiae
Anopheles
Freund's Adjuvant
Antibodies
Escherichia coli Proteins
Culicidae
Serum
Germ Cells
Epitopes
Immunoglobulin G
Monoclonal Antibodies

ASJC Scopus subject areas

  • Immunology
  • Microbiology
  • Parasitology
  • Infectious Diseases

Cite this

Potent malaria transmission-blocking antibody responses elicited by Plasmodium falciparum Pfs25 expressed in Escherichia coli after successful protein refolding. / Kumar, Rajesh; Angov, Evelina; Kumar, Nirbhay.

In: Infection and Immunity, Vol. 82, No. 4, 2014, p. 1453-1459.

Research output: Contribution to journalArticle

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