National Institute of Genetic Engineering and Biotechnology (NIGEB).
Daru. 2010;18(1):64-8.
Heat Shock Protein 90 (Hsp90) is typically the most abundant chaperone in the eukaryotic cell cytoplasm, and its expression is essential for loading immunogenic peptides onto major histocompatibility complex molecules for presentation to T-cells. Therefore, it may act as a good candidate as an adjuvant molecule in vaccine technology.
Initially the human Hsp90β gene was cloned into the heat inducible expression vector pGP1-2 and then the recombinant protein was isolated by ion exchange chromatography. After intradermal injection of confirmed purified band of protein to rabbits and isolation of the serum IgG antibody, for its affinity purification, the rabbit's purified Hsp90 specific IgG was coupled to the cyanogen bromide-activated Sepharose 4B.
The recovery of the purified protein of interest by affinity chromatography was 50%.
This research enabled purification of human heat shock protein by a laboratory prepared column chromatography.
热休克蛋白 90（Hsp90）通常是真核细胞质中最丰富的伴侣蛋白，其表达对于将免疫原性肽加载到主要组织相容性复合物分子上以呈递给 T 细胞是必不可少的。因此，它可能是疫苗技术中作为佐剂分子的良好候选物。
最初将人 Hsp90β 基因克隆到热诱导表达载体 pGP1-2 中，然后通过离子交换色谱法分离重组蛋白。在向兔子皮内注射确认的纯化蛋白条带并分离血清 IgG 抗体后，为了进行亲和纯化，将兔的纯化 Hsp90 特异性 IgG 与氰基溴化活化的 Sepharose 4B 偶联。
亲和层析法回收的纯化目的蛋白为 50%。
本研究通过实验室制备的柱层析法实现了人热休克蛋白的纯化。
