Recombinant expression and characterization of CD2-binding domain of Macaca mulatta lymphocyte function-associated antigen 3 in Pichia pastoris.
- Author:
Jian ZHU
;
Shengyun ZHU
;
Hao YANG
;
Xiaofeng LU
;
Lin WAN
- Publication Type:Journal Article
- MeSH:
Animals;
CD58 Antigens;
biosynthesis;
Humans;
Immunoglobulin G;
Lymphocyte Activation;
Macaca mulatta;
Pichia;
Plasmids;
Protein Interaction Domains and Motifs;
Recombinant Fusion Proteins;
biosynthesis;
T-Lymphocytes
- From:
Journal of Biomedical Engineering
2015;32(1):120-125
- CountryChina
- Language:Chinese
-
Abstract:
Human lymphocyte function-associated antigen 3 (hLFA3) has been identified as an important T cell accessory molecule. Rhesus monkeys (Macaca mulatta) have been widely used as animal models for human immune disorders. Due to the species-specificity of immune system, it is necessary to study M. mulatta LFA3 (mmLFA3). In this study, the gene encoding mmLFA3 CD2-binding domain (mmLFA3Sh) was amplified by polymerase chain reaction (PCR) and genetically fused to human IgG1 Fc fragment in pPIC9K to construct the expression plasmid pPIC9K-mmLFA3Sh-Ig. Approximately 3-4 mg mmLFA3Sh-Ig protein was recovered from 1 L of inductive media, and mmLFA3Sh-Ig produced by the P. pastoris can bind to the CD2 positive cells, and suppress the monkey and human lymphocytes proliferation induced by Con A and alloantigen in a dose-dependent manner. These results suggested that mmLFA3Sh-Ig might be used as a novel tool for pathogenesis and experimental immunotherapy of Rhesus monkey immune disorders.
