HDL-ApoA1 plays a pivotal role in the prevention of atherosclerosis and cardiovascular diseases. ApoA1 purification from blood plasma has always remained tedious, involving multiple steps, large volumes of plasma and substantial loss in the final yield of pure ApoA1. In this study, a two-step method has been developed and optimized for the purification of ApoA1 from plasma. Plasma was first subjected to 60% ammonium sulphate (NH₄)₂SO₄ precipitation and subsequently, ApoA1 was recovered using mixed mode chromatographic sorbent, HEA HyperCel™. ApoA1 was found to be enriched in 60% (NH₄)₂SO₄ supernatant that was dialyzed and injected onto HEA sorbent with 50 mM phosphate buffer pH 7.4. The bound proteins were eluted by decreasing the pH in step-gradient from pH 7.4 to pH 4.0 and subsequently to pH 3.5 using 50 mM sodium acetate buffer. Gel electrophoresis showed elution of homogeneous apoA1 at pH 3.5, with purity and yield of 63%. An interesting feature of this approach is that the purified ApoA1 was monomeric with a mass of 28,079.30 Da as confirmed by MS analysis. This simple and efficient method of purification of apoA1 serves as an alternative method which can be combined with traditional approaches and has a great potential for biochemical and clinical studies.

HDL-ApoA1在预防动脉粥样硬化和心血管疾病中起关键作用。从血浆中纯化ApoA1一直很繁琐，涉及多个步骤，大量血浆以及纯ApoA1最终产量的重大损失。在这项研究中，已经开发了一种两步法并对其进行了优化，以从血浆中纯化ApoA1。首先使血浆经受60％硫酸铵（NH ₄）₂ SO ₄沉淀，然后使用混合模式色谱吸附剂HEA HyperCel™回收ApoA1。发现ApoA1富含60％（NH ₄）₂ SO ₄透析的上清液注入具有50 mM磷酸盐缓冲液pH 7.4的HEA吸附剂中。通过使用50 mM乙酸钠缓冲液将pH逐步从pH 7.4降低到pH 4.0，然后降低到pH 3.5来洗脱结合的蛋白。凝胶电泳显示在pH 3.5下洗脱均质的载脂蛋白A1，纯度和产率为63％。该方法的一个有趣特征是纯化的ApoA1是单体，质量为28,079.30 Da，通过MS分析确认。这种简单而有效的纯化apoA1的方法可作为替代方法，可与传统方法结合使用，在生化和临床研究中具有巨大潜力。