Histidine affinity tags affect MSP1(42) structural stability and immunodominance in mice

Biotechnol J. 2012 Jan;7(1):133-47. doi: 10.1002/biot.201100331. Epub 2011 Dec 28.

Abstract

Inclusion of affinity tags has greatly facilitated process development for protein antigens, primarily for their recovery from complex mixtures. Although generally viewed as supportive of product development, affinity tags may have unintended consequences on protein solubility, susceptibility to aggregation, and immunogenicity. Merozoite surface protein 1 (MSP1), an erythrocytic stage protein of Plasmodium falciparum and a candidate malaria vaccine, was used to evaluate the impact of a metal ion affinity-tag on both protein structure and the induction of immunity. To this end, codon harmonized gene sequences from the P. falciparum MSP1(42) of FVO and 3D7 parasites were cloned and purified with and without a histidine (His) tag. We report on the influence of His-affinity tags on protein expression levels, solubility, secondary structure, thermal denaturation, aggregation and the impact on humoral and cellular immune responses in mice. While the overall immunogenicity induced by His-tagged MSP1(42) proteins is greater, the fine specificity of the humoral and cellular immune responses is altered relative to anti-parasitic antibody activity and the breadth of T-cell responses. Thus, the usefulness of protein tags may be outweighed by their potential impact on structure and function, stressing the need for caution in their use. See accompanying commentary by Randolph DOI: 10.1002/biot.201100459.

Publication types

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

MeSH terms

  • Affinity Labels / chemistry*
  • Animals
  • Antibodies, Protozoan / chemistry
  • Antibodies, Protozoan / genetics
  • Antibodies, Protozoan / immunology
  • Histidine / chemistry*
  • Histidine / immunology*
  • Immunity, Cellular / genetics
  • Immunity, Cellular / immunology
  • Malaria Vaccines / chemistry
  • Malaria Vaccines / genetics
  • Malaria Vaccines / immunology
  • Merozoite Surface Protein 1 / chemistry*
  • Merozoite Surface Protein 1 / genetics
  • Merozoite Surface Protein 1 / immunology*
  • Mice
  • Mice, Inbred BALB C
  • Plasmodium falciparum / genetics
  • Plasmodium falciparum / metabolism
  • Protein Denaturation
  • Protein Folding
  • Protein Structure, Secondary
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Solubility
  • T-Lymphocytes / immunology

Substances

  • Affinity Labels
  • Antibodies, Protozoan
  • Malaria Vaccines
  • Merozoite Surface Protein 1
  • Recombinant Proteins
  • Histidine