Rapid purification method maintaining cystatin C antibody-latex covalently coupled compound activity

A technology of covalent coupling and purification method, applied in the field of separation and purification, can solve the problems of small scale, unsuitable for industrial production, etc., and achieve the effect of short time and high reactivity

Inactive Publication Date: 2016-12-07
WENZHOU PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a purification method that maintains the activity of the cystatin C antibody-latex covalent coupling complex, aiming at solving the problem of the existing centrifugation method when purifying the cystatin C antibody-latex covalent coupling complex The problem of small scale and not suitable for industrial production

Method used

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  • Rapid purification method maintaining cystatin C antibody-latex covalently coupled compound activity
  • Rapid purification method maintaining cystatin C antibody-latex covalently coupled compound activity

Examples

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Embodiment 1

[0039] Purification of 600ml Cystatin C Antibody-Latex Covalently Covalently Conjugated Complex Solution; Prepare Cystatin C Antibody-Latex Covalently Conjugated Complex Crude Solution with Polystyrene Latex with a Diameter of 80nm, Follow the steps below purification:

[0040] 1) Transfer 600ml of crude cystatin C antibody-latex covalent coupling complex solution to be purified to the sample container of the tangential flow filtration system, set the ultrafiltration membrane inside the hollow fiber membrane filter, and the ultrafiltration membrane It is a modified polyethersulfone membrane with a molecular weight cut-off of 500KD and a membrane area of ​​115cm 2 , set the target pump speed to 30ml / min, and the shear force to 2000s -1 , close the filtrate clamp of the filtration circuit, first adjust the pump speed to half of the target flow rate (15ml / min), turn on the peristaltic pump, and fill the entire filtration pipeline with the coarse solution of Cystatin C antibody-l...

Embodiment 2

[0048] Purification of 4L cystatin C antibody-latex covalently coupled complex crude solution; use polystyrene latex with a diameter of 100nm to prepare cystatin C antibody-latex covalently coupled complex crude solution, proceed as follows purification:

[0049] 1) Transfer 4L of crude cystatin C antibody-latex covalent coupling complex solution to be purified to the sample container of the tangential flow filtration system, set the ultrafiltration membrane inside the hollow fiber membrane filter, and the ultrafiltration membrane It is a modified polyethersulfone membrane with a molecular weight cut-off of 500KD and a membrane area of ​​790cm 2 , set the target pump speed to 420ml / min, and the shear force to 2000s -1. Close the filtrate clamp of the filtration circuit, first adjust the pump speed to half of the target flow rate (210ml / min), turn on the peristaltic pump, and fill the entire filtration pipeline with the coarse solution of Cystatin C antibody-latex covalent co...

Embodiment 3

[0058] Purification of 10L crude solution of cystatin C antibody-latex covalently coupled complex; use polystyrene latex with a diameter of 120nm to prepare the crude solution of cystatin C antibody-latex covalently coupled complex, proceed as follows purification:

[0059] 1) Transfer 10L of crude cystatin C antibody-latex covalent coupling complex solution to be purified to the sample container of the tangential flow filtration system, set the ultrafiltration membrane inside the hollow fiber membrane filter, and the ultrafiltration membrane It is a modified polyethersulfone membrane with a molecular weight cut-off of 500KD and a membrane area of ​​2600cm 2 , set the target pump speed to 1.15L / min, and the shear force to 2000s -1 . Close the filtrate clamp of the filtration circuit, first adjust the pump speed to half of the target flow rate (575ml / min), turn on the peristaltic pump, and fill the entire filtration pipeline with the coarse solution of Cystatin C antibody-lat...

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Abstract

The invention discloses a purification method maintaining cystatin C antibody-latex covalently coupled compound activity. The method consists of: transferring a to-be-purified cystatin C antibody-latex covalently coupled compound crude solution to a tangential flow filtration system, setting the shear force, arranging an ultrafiltration membrane in a hollow fiber membrane filter, wherein the ultrafiltration membrane is a modified polyethersulfone membrane with a molecular weight cut-off of 500KD, setting the shearing force at 2000s<-1>, a corresponding membrane area of 115cm<2>-2600cm<2> and a pump speed of 30ml/min-1150ml/min; concentrating the solution within 60min by 5 times to obtain a crude concentrated cystatin C antibody-latex covalently coupled compound solution; and within 60min using a buffer solution to elute the crude concentrated cystatin C antibody-latex covalently coupled compound solution by 5 times. The method provided by the invention needs short time, the purified antibody-latex compound has high reaction activity, is not only suitable for small-scale preparation in laboratories, but also suitable for industrial large-scale production.

Description

technical field [0001] The invention belongs to the technical field of separation and purification, and in particular relates to a purification method for maintaining the activity of a cystatin C antibody-latex covalent coupling complex. Background technique [0002] Cystatin C (CysC), also known as cystatin C, is a low-molecular-weight protein discovered by scientists such as Simonsen in Sweden in the mid-1980s. Research data have shown that in the diagnosis of renal insufficiency, liver cirrhosis, tumors and other diseases, CysC can well replace serum creatinine and creatinine elimination rate, and become a new reflection of glomerular filtration rate ( Ideal marker for GFR). Nano-latex microsphere-enhanced transmission immunoturbidimetry (PETIA) is the latest method for the clinical determination of cystatin C. During the preparation of the detection reagent of this method, the activity of the cystatin C antibody-latex covalently coupled complex is maintained Rapid puri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/38C07K1/34
Inventor 潘利琴许锴陈益川张德亭连国军
Owner WENZHOU PEOPLES HOSPITAL
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