Abstract：
Antibodies from sera of a multiple sclerosis (MS) patient subpopulation preferentially recognize the hyperglucosylated adhesin protein HMW1ct(Glc) of the pathogen Haemophilus influenzae. This protein is the first example of an N-glucosylated native antigen candidate, potentially triggering pathogenic antibodies in MS. Specific antibodies in patients\' sera can be isolated exploiting their biospecific interaction with antigens by affinity chromatography. Herein, the proteins HMW1ct and HMW1ct(Glc) were first immobilized on appropriately functionalized supports and further used to purify antibodies directly from MS patients sera. We describe a protocol to obtain an antibody fraction specifically recognizing the glusosylated residues on the HMW1ct(Glc) adhesin protein depleting antibodies to the unglucosylated HMW1ct sequence. Different elution solutions have been tested to recover the purified antibody fraction, strongly bound to the immobilized HMW1ct(Glc) adhesin protein.
摘要：
来自多发性硬化症（MS）患者亚群血清的抗体优先识别病原体流感嗜血杆菌的高糖基化粘附素蛋白HMW1ct（Glc）。该蛋白是N-糖基化天然候选抗原的第一个例子，可能触发MS中的致病性抗体。患者血清中的特异性抗体可以通过亲和层析利用其与抗原的生物特异性相互作用来分离。在这里，首先将蛋白质HMW1ct和HMW1ct（Glc）固定在适当功能化的支持物上，并进一步用于直接从MS患者血清中纯化抗体。我们描述了获得抗体级分的方案，该抗体级分特异性识别HMW1ct（Glc）粘附素蛋白消耗抗体上的未糖基化HMW1ct序列的糖基化残基。已经测试了不同的洗脱溶液以回收纯化的抗体级分，与固定的HMW1ct（Glc）粘附素蛋白强烈结合。
