The aim of this study was to establish an assessment method for determiningα-Gal(α-1, 3-galactosyle) epitopes contained in animal tissue or animal tissue-derived biological materials with ELISA inhibition assay. Firstly, a 96 well plate was coated with Galα-1, 3-Gal/bovine serum albumin (BSA) as a solid phase antigen and meanwhile, the anti-α-Gal M86 was used to react withα-Gal antigens which contained in the test materials. Then, the residual antibodies (M86) in the supernatant of M86-Gal reaction mixture were measured using ELISA inhibition assay by theα-Gal coating plate. The inhibition curve of the ELISA inhibition assay, the R2=0.999, was well established. Checking using bothα-Gal positive materials (rat liver tissues) andα-Gal negative materials (human placenta tissues) showed a good sensitivity and specificity. Based on the presently established method, theα-Gal expression profile of rat tissues, decellular animal tissue-derived biological materials and porcine dermal before and after decellular treatment were determined. The M86 ELISA inhibition assay method, which can quantitatively determine theα-Gal antigens contained in animal tissues or animal tissue-derived biomaterials, was refined. This M86 specific antibody based-ELISA inhibition assay established in the present study has good sensitivity and specificity, and could be a useful method for determining remnantα-1, 3Gal antigens in animal tissue-derived biomaterials.
ObjectiveTo investigate the distribution and content of endogenous salusin-βin septic rats. MethodsThirty-six SPF male SD rats were randomly divided into sham operation group (n=9) and septic model group (n=27).Only the cecum was turn in the sham operation group and the septic model was made by the cecal ligation and puncture (CLP) in the septic model group.The rats were sacrificed at 6 h, 12 h, and 24 h after modeling in the septic model group.The contents of salusin-βin the tissues of spleen, stomach, small intestine, hypothalamus, and serum specimens were detected by enzyme-linked immunosorbent assay. Results①The salusin-βendogenously generated in the rat tissues including the spleen, stomach, small intestine, hypothalamus, and serum.The content of salusin-βin the spleen tissue was higher than that in the other tissues (P < 0.05).②The contents of salusin-βin the spleen, stomach, small intestine tissues together with the serum increased significantly at 6 h after CLP as compared with the sham operation group (P < 0.05).The contents of salusin-βin the spleen tissue and serum were peaked at 12 h, in the small intestine tissue reached the summit at 24 h.While, the content of salusin-βhad no significant fluctuation in the stomach tissue.The content of salusin-βbegan to increase at 6 h in the hypothalamus tissue, and significantly increased at 12 h after CLP (P < 0.05). ConclusionThe time-dependent change of salusin-βin sepsis rats suggests that salusin-βmight be involved with the pathogenesis of sepsis.