High-yield production of recombinant platelet factor 4 by harnessing and honing the gram-negative bacterial secretory apparatus.

ABSTRACT: Platelet factor 4 is a cytokine released into the bloodstream by activated platelets where it plays a pivotal role in etiology and diagnosis of heparin-induced thrombocytopenia. Therefore, a sustainable source of recombinant PF4 with structural and functional similarity to its native form is urgently needed to be used in diagnostic procedures. To this end, a three-in-one primary construct was designed from which three secondary constructs can be derived each capable of employing either type I, type II secretory or cytoplasmic pathways. Protein expression and secretion were performed in Escherichia coli BL21 (DE3) and confirmed by SDS-PAGE and Western blotting. To further enhance protein secretion, the effect of several controllable chemical factors including IPTG, Triton X-100, sucrose, an

SUBMITTER: Ataei S 

PROVIDER: S-EPMC7205247 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature