Method for separating and purifying collagen for biopharmaceutical use from sharkskin

Abstract

A method for obtaining collagen for biomedical use from shark skin through separation and purification adopts the raw material of the shark skin. The method has the following steps: steeping the shark skin in low-concentration phosphate buffer solution with a PH value of 7.4-7.6 to flush away squamae and residual flesh, chopping the shark skin in small pieces, and steeping and flushing the shark skin with deionized water for 3-5 times; steeping the clean shark skin in medium-concentration phosphate buffer solution which is five folds of the raw material amount with a PH value of 7.4-7.6 for 70-75 hours, then stirring the pulp evenly for 15 minutes with a temperature of 4 DEG C and a rotating speed of 10000rpm, and centrifuging the pulp for 15 minutes and taking supernatant; depositing and repeating the former step, mixing supernatant, adding ammonium sulphate to a saturation degree of 80 percent, a temperature of 4 DEG C and a rotating speed of 8000rpm, and centrifuging for 15 minutes and retaining deposit; dissolving the deposit in medium-concentration phosphate buffer solution with a PH value of 7.0; chromatographically fractionating the solution with G-75, 100, 150 glucan, collecting albumen digesting liquid with a temperature of 4 DEG C and a rotating speed of 10000rpm, centrifuging for 15 minutes, concentrating supernatant, and obtaining the collagen for biomedical use after freeze drying.

Description

Technical field: [0001] The invention relates to a method for separating and purifying collagen for biomedicine, in particular to a method for separating and purifying collagen for biomedicine from shark skin. Background technique: [0002] Collagen or collagen is a biological polymer synthesized by animal cells. It is widely found in animal bones, feet, cartilage, skin and other connective tissues, accounting for about 1 / 3 of the total mammalian protein. The extracellular matrix component, collagen, has good biological properties and can be used as a tissue support, which has a significant impact on the normal function of cells, tissues and even organs and has a significant impact on the repair of trauma. The utilization of collagen at home and abroad is mainly divided into two categories: one is to focus on the unique macroscopic physical properties of collagen, namely fiber properties, which are used for photographic paper negatives, textiles, papermaking, etc.; the other...

AI Technical Summary

Problems solved by technology

Regarding the extraction and purification of collagen, although the methods are different, most of them are extracted from the skin of small fish such as squid and cod and livestock. Small fish itself contains less collagen and the raw materials are limited. The extracted protein has low solubility, and the use of cattle as raw materials is not easily accepted by many trade organizations in the world. The current steps of extracting collagen include extraction, separation and purification. Extraction is divided into acid method, alkali method, neutral salt method and Enzyme method, etc., the process is loaded down with trivial details, and protein is also prepared by genetic engineering method, but increases cost again at the same time, because these defects that above-mentioned these separation techniques exist, make the collagen production rate, output, quality that make is not high, so that The high price of collagen for biomedical use finally greatly affects the wide application of collagen. Therefore, reducing the cost of collagen purification and improving protein quality are the key