Cabrera-Artiles Y, Veitia I, Barceló MT, Martínez D, Hernández I, Leal V, Armas R, Rubí JA

Language: English
References: 0
Page: 1311-1317
PDF size: 1243.33 Kb.

ABSTRACT

The multi-epitope polypeptide TAB1 representing sequences of the gp120 HIV protein is one of the antigens included in the UMELISA® HIV 1+2 RECOMBINANT diagnostic kit to detect antibodies against HIV. This test is produced by the Immunoassay Center and it has been used in Cuba both for screening HIV infected patients and checking blood quality in blood Banks. Originally the protein was expressed in a vector with the tryptophan promoter and was purified by reverse phase HPLC, which is an expensive and low yielding process. In this study, the expression vector was redesigned, using new cloning and purification techniques, by including a gene modification that added a hexahistidine tag to TAB1, allowing for its purification with immobilized metal affinity chromatography. The replacement of the tryptophan promoter regulatory system by the hybrid ptrc promoter offered better control and an expression level of about 20%. This new process enabled the production of the TAB1-His protein with the appropriate purity profile and antigenic properties for its binding to HIV antibodies, while meeting the requirements for its use in the diagnostic kit UMELISA® HIV 1+2 RECOMBINANT.