Acapsular type staphylococcus aureus extracellular polysaccharide-protein conjugate and preparation method thereof

A staphylococcus and extracellular polysaccharide technology, applied in the field of non-capsulated Staphylococcus aureus exopolysaccharide-protein conjugates and its preparation, can solve the problems of difficult batch preparation, complicated purification process, and inconsistent coupling reaction system Stability and other issues, to achieve the effect of increasing the level of humoral immunity, stable coupling reaction conditions, and stable reaction conditions

Inactive Publication Date: 2011-07-13
新疆维吾尔自治区畜牧科学院兽医研究所
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

When using this method for purification and coupling, the purification process is relatively complicated, the coupling reaction system is unstable, and it is not easy to prepare in batches

Method used

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  • Acapsular type staphylococcus aureus extracellular polysaccharide-protein conjugate and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 A non-capsulated Staphylococcus aureus exopolysaccharide-protein conjugate, composed of the non-capsulated Staphylococcus aureus exopolysaccharide and bovine serum albumin under the action of coupling reagents EDAC and Sulfo-NHS A non-capsulated Staphylococcus aureus exopolysaccharide-bovine serum albumin conjugate prepared by covalent coupling.

[0037] The preparation method of non-capsulated staphylococcus aureus polysaccharide-bovine serum albumin conjugate prepared by EDAC zero-distance condensation method is as follows:

[0038] (1) Isolation and purification of non-encapsulated Staphylococcus aureus exopolysaccharide

[0039] ① Inoculate Staphylococcus aureus strains in Columbia liquid medium (containing 2% sodium chloride), shake culture at 170 rpm at 37°C for 22 to 24 hours, centrifuge at 5000 rpm for 30 minutes to collect bacterial precipitates, weigh Freeze at -75°C.

[0040] ② Suspend the collected bacterial pellet in 50mmol / L Tris containing 2mm...

Embodiment 2

[0052] Example 2 A non-encapsulated Staphylococcus aureus exopolysaccharide-protein conjugate, in which the exopolysaccharide of non-encapsulated Staphylococcus aureus is covalently bound to bovine serum protein under the action of coupling reagents EDAC and ADH A non-capsulated staphylococcus aureus exopolysaccharide-bovine serum protein conjugate prepared by coupling.

[0053] The improved ADH bridge method prepares the preparation method of non-capsulated Staphylococcus aureus exopolysaccharide-bovine serum albumin conjugate, specifically as follows:

[0054] (1) Isolation and purification of non-encapsulated Staphylococcus aureus exopolysaccharide, the method is the same as above.

[0055] (2) Make the purified polysaccharide into a 10mg / mL solution with 100mmol / L MES buffer with a pH value of 5.6, take 1.0mL of the solution, add ADH and EDAC to the final concentrations of 0.36mol / L and 0.015mol / L respectively , mix well, and react at room temperature to form derivatized ...

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Abstract

The invention relates to an acapsular type staphylococcus aureus extracellular polysaccharide-protein conjugate and a preparation method thereof. The acapsular type staphylococcus aureus extracellular polysaccharide-protein conjugate is prepared by performing covalent coupling on extracellular polysaccharide of acapsular type staphylococcus aureus and protein under the action of a coupling reagent. The preparation method is an antidiuretic hormone (ADH) bridge method or an l-ethyl-3(3-diaminopropyol)-carbodiimide (EDAC) zero-distance crosslinking method. The preparation of the acapsular type staphylococcus aureus extracellular polysaccharide-protein conjugate provides a strong technical support for development of a vaccine for preventing staphylococcus aureus.

Description

technical field [0001] The invention relates to an uncapsulated staphylococcus aureus exopolysaccharide-protein conjugate and a preparation method thereof. Background technique [0002] Staphylococcus aureus ( Staphylococcus aureus ) is one of the main pathogens of cow mastitis and iatrogenic infection. Current research shows that there are 11 serotypes of Staphylococcus aureus, and most of the Staphylococcus aureus isolated from clinical cases are capsular polysaccharide type 5 (type 5 capsular polysaccharide, CP5), capsular polysaccharide type 8 (type 8 capsular polysaccharide, CP8) and non-capsulated type 336 (type 336 polysaccharides, 336PS), and the main circulating serotypes are different in different countries and regions. The typing results of Verdier I (2007) on 195 iatrogenic Staphylococcus aureus showed that CP5 type accounted for 42%, CP8 type accounted for 45%, and 336 type accounted for 13%. The typing results of Melles D C (2008) on 162 strains of methicill...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/31C07K1/107
Inventor 王治才吴建勇杨学云王登峰
Owner 新疆维吾尔自治区畜牧科学院兽医研究所
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