The current study deals with the purification and characterization of non-enzymatic glycoprotein (NEGp) from flax seed buffer extract. Sephadex G-100 and DEAE-A25 column chromatography techniques were employed to isolate NEGp. NEGp showed single sharp band at 29 kDa region on 10% SDS-PAGE, and under reduced and non-reduced conditions revealed its monomeric nature. Besides, NEGp taken up the PAS stain at 29 kDa region reveals the presence of carbohydrate moiety. Purity of NEGp was adjudged by RP-HPLC, as it revealed a single sharp peak at the retention time of 3.4 min. The exact molecular mass of NEGp was found to be 26 kDa which was confirmed by MALDI-TOF. Circular di-chromism spectra of NEGp showed 12.0% α-helix, 24.3% α-helix turn and 63.7% random coils without beta pleated sheets. NEGp was found to exhibit anticoagulant activity by extending clotting time of both platelet rich plasma and platelet poor plasma from control 240 s to 1800 s and 280 s to 2100 s respectively at the concentration of 8 μg. NEGp inhibited the agonists such as ADP, epinephrine and arachidonic acid induced platelet aggregation in washed platelets. The percentage of inhibition was found to be 70%, 80% and 60% respectively. While, it did not interfere in thrombin, PAF and collagen induced platelet aggregation. NEGp did not hydrolyse RBC membrane, devoid of haemorrhagic and edema inducing properties in experimental mice.

目前的研究涉及从亚麻籽缓冲液提取物中纯化和表征非酶糖蛋白（NEGp）。用Sephadex G-100和DEAE-A25柱色谱技术分离NEGp。NEGp在10％SDS-PAGE上在29 kDa区域显示单个锐带，并且在还原和非还原条件下均显示出其单体性质。此外，NEGp在29 kDa区域吸收了PAS染色，表明存在碳水化合物部分。RP-HPLC判断NEGp的纯度，因为它在保留时间3.4分钟时显示出一个尖锐的峰。发现NEGp的确切分子量为26kDa，这由MALDI-TOF证实。NEGp的圆形双色光谱显示12.0％的α-螺旋，24.3％的α-螺旋转角和63.7％的无规则褶皱卷，无β折叠片。发现NEGp通过在浓度为8μg的情况下分别将富血小板血浆和贫血小板血浆的凝血时间分别从对照组的240 s延长至1800 s和将280 s延长至2100 s，从而表现出抗凝活性。NEGp抑制了激动剂，例如ADP，肾上腺素和花生四烯酸诱导的血小板聚集。发现抑制百分比分别为70％，80％和60％。同时，它不干扰凝血酶，PAF和胶原蛋白诱导的血小板聚集。NEGp没有水解RBC膜，缺乏实验小鼠的出血和水肿诱导特性。肾上腺素和花生四烯酸诱导血小板在洗涤过的血小板中聚集。发现抑制百分比分别为70％，80％和60％。同时，它不干扰凝血酶，PAF和胶原蛋白诱导的血小板聚集。NEGp没有水解RBC膜，缺乏实验小鼠的出血和水肿诱导特性。肾上腺素和花生四烯酸诱导血小板在洗涤过的血小板中聚集。发现抑制百分比分别为70％，80％和60％。同时，它不干扰凝血酶，PAF和胶原蛋白诱导的血小板聚集。NEGp没有水解RBC膜，缺乏实验小鼠的出血和水肿诱导特性。