A novel adjuvant-free H fusion system for the production of recombinant immunogens in Escherichia coli: Its application to a 12 kDa antigen from Cryptosporidium parvum.
The production of recombinant antigens in Escherichia coli and specific polyclonal antibodies for diagnosis and therapy is still a challenge for world-wide researchers. Several different strategies have been explored to improve both antigen and antibody production, all of them depending on a successful expression and immunogenicity of the antigen. Gene fusion technology attempted to address these challenges: fusion partners have been applied to optimize recombinant antigen production in E. coli, and to increase protein immunogenicity. Taking a 12-kDa surface adhesion antigen from Cryptosporidium parvum (CP12) by example, the novel H fusion partner was presented in this work as an attractive option for the development of recombinant immunogens and its adjuvant-free immunization. The H tag (of only 1 kDa) efficiently triggered a CP12-specific immune response, and it also improved the immunization procedure without requiring co-administration of adjuvants. Moreover, polyclonal antibodies raised against the HCP12 fusion antigen detected native antigen structures displayed on the surface of C. parvum oocysts. The H tag proved to be an advanced strategy and promising technology for the diagnosis and therapy of C. parvum infections in animals and humans, allowing a rapid and simple recombinant production of the CP12 antigen.
Institute for Biotechnology and Bioengineering; Centre of Biological Engineering; Universidade do Minho, Campus de Gualtar; Braga, Portugal; Instituto Nacional de Saúde Dr Ricardo Jorge; Porto, Portugal; Escola Superior Agrária de Coimbra; Coimbra, Portugal; Instituto de Ciências e Tecnologias Agrárias e Agroalimentares da Universidade do Porto; Porto Portugal; Hitag Biotechnology, Lda.; Biocant-Parque Tecnológico de Cantanhede; Cantanhede, Portugal.
Sofia J
SJ
Costa
Instituto Nacional de Saúde Dr Ricardo Jorge; Porto, Portugal; Escola Superior Agrária de Coimbra; Coimbra, Portugal.
Pedro
P
Silva
Instituto de Ciências e Tecnologias Agrárias e Agroalimentares da Universidade do Porto; Porto Portugal; Hitag Biotechnology, Lda.; Biocant-Parque Tecnológico de Cantanhede; Cantanhede, Portugal.
André
A
Almeida
Escola Superior Agrária de Coimbra; Coimbra, Portugal.
Antónia
A
Conceição
Institute for Biotechnology and Bioengineering; Centre of Biological Engineering; Universidade do Minho, Campus de Gualtar; Braga, Portugal.
Lucília
L
Domingues
Instituto Nacional de Saúde Dr Ricardo Jorge; Porto, Portugal.
António
A
Castro
D000595:Amino Acid Sequence; D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D000953:Antigens, Protozoan / Q000096:biosynthesis*; D001483:Base Sequence; D016785:Cryptosporidium parvum / Q000235:genetics; D016054:DNA, Protozoan / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D050939:Gene Fusion; D051379:Mice; D008969:Molecular Sequence Data; D033661:Oocysts / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis*
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D016054:DNA, Protozoan; D011994:Recombinant Proteins
CP12; Cryptosporidium; antibody production; free-adjuvant immunization; immunogens; novel fusion partner