Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Human NT-proBNP specificity recombinant sheep monoclonal antibody and preparation method and application thereof

A technology for cloning antibodies, nt-probnp, which is applied in the field of recombinant goat monoclonal antibodies and its preparation, can solve the problems of high storage cost of hybridoma cells, complex screening of hybridomas, and differences between batches of monoclonal antibodies, etc.

Active Publication Date: 2019-04-26
CHONGQING TANSHENG TECH
View PDF7 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the preparation process of monoclonal antibodies, there are situations such as complex screening of hybridomas, high storage costs of hybridoma cells, and the need to feed experimental mice.
At present, there are different degrees of batch-to-batch differences in the preparation of monoclonal antibodies by the mouse ascites induction method, which brings greater workload to product quality control.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Human NT-proBNP specificity recombinant sheep monoclonal antibody and preparation method and application thereof
  • Human NT-proBNP specificity recombinant sheep monoclonal antibody and preparation method and application thereof
  • Human NT-proBNP specificity recombinant sheep monoclonal antibody and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0028] According to a specific embodiment of the present invention, the human NT-proBNP-specific recombinant goat monoclonal antibody consists of the goat single-chain antibody scFv and the Fc segment of human IgG.

[0029] According to the present invention, the selection of the Fc segment of human IgG is not particularly limited, and may be various Fc segments of human IgG known in the prior art. According to a specific embodiment of the present invention, the amino acid sequence of the Fc segment of human IgG is shown in SEQ ID NO:3.

[0030] The amino acid sequence of the Fc segment of human IgG shown in SEQ ID NO: 3 is as follows:

[0031]ESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK(SEQ ID NO:3)。

[0032] In the second aspect, the present invention also provides the gene encoding the goat ...

Embodiment 1

[0081] This example is used to illustrate the preparation of human NT-proBNP-specific recombinant goat monoclonal antibody

[0082] (1) Preparation of immunogen

[0083] Apply online B cell epitope prediction software (http: / / tools.immunepitope.org / bcell / ) to analyze potential B cell epitope dominant antigen epitopes in human NT-proBNP, and select 2 peptides as immunogens: peptide 1 (amino acid sequence is HPLGSPGSASDLETSG, SEQ ID NO: 5) and peptide 2 (amino acid sequence is QRNHLQGKLSELQVE, SEQ ID NO: 6). Chemically synthesized by Gill Biochemical (Shanghai) Co., Ltd. and coupled with bovine serum albumin (BSA) through the N-terminal amino group.

[0084] (2) Preparation of immune antibody library

[0085] ①The synthetic epitope peptide-BSA conjugate was emulsified with the same amount of Freund's complete adjuvant (Sigma Company) by double-syringe cross-push method, and an adult Boer sheep was administered to an adult Boer sheep by subcutaneous multi-point injection on bot...

Embodiment 2

[0103] This example is used to illustrate the method for detecting human NT-proBNP in samples by double-antibody sandwich ELISA

[0104] (1) Human NT-proBNP-specific goat monoclonal antibody coated microtiter plate

[0105] ① Take NT-proBNP-specific goat monoclonal antibody coating solution (NaHCO 3 2.93g, Na 2 CO 3 1.59g, dissolved in double distilled water, total volume 1L, pH 9.6, analytical grade) diluted to 5μg / mL, prepare 24mL for each ELISA plate; add to 96-well ELISA plate, 200μL per well; affix a seal, 4 °C overnight.

[0106] ② Shake off the antigen, wash the ELISA plate 5 times with clean tap water, tap it on the absorbent paper for a few times after washing until there is no obvious liquid residue; use the washing solution (Na 2 HPO 4 0.14g, Tween-20 1mL, double-distilled water to 1L, analytically pure) to prepare 5% bovine serum albumin (purchased from Beyentian Biotechnology), add 300 μL to each well, and place at room temperature for 2 hours; shake off t...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to the technical field of biological detection, and particularly relates to a human brain sodium peptide original N-terminal peptide (NT-proBNP) specificity recombinant sheep monoclonal antibody and a preparation method and application thereof. The human NT-proBNP specificity recombinant sheep monoclonal antibody comprises a sheep single-chain antibody svFv and an Fc segmentof human IgG, wherein an amino acid sequence of the sheep single-chain antibody svFv is as shown in SEQ ID NO: 1. The human NT-proBNP specificity recombinant sheep monoclonal antibody is capable of realizing low-cost, high-efficient and high-quality large-scale preparation of the recombinant antibody, and the obtained antibody has high affinity to human NT-proBNP, so a core raw material foundationis established for preparing a human NT-proBNP immunodetection kit with more super quality and competitive price.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a recombinant goat monoclonal antibody specific to human brain natriuretic peptide N-terminal peptide (NT-proBNP) and its preparation method and application. Background technique [0002] Cardiovascular disease is an important disease that seriously threatens human life and health. Among them, heart failure (Heart Failure, HF) is a pathological syndrome in cardiovascular diseases that seriously endanger human health. disease, diabetes, etc.) to left ventricular hypertrophy, myocardial cell dysfunction, coronary heart disease, and finally develops into heart failure. Since the incidence of coronary heart disease and hypertension is still on the rise in my country, and the population aging trend is obvious, heart failure is becoming an important public health problem in the cardiovascular field in my country, and its early diagnosis and early treatment are extremely i...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/26C12N15/13G01N33/74G01N33/577
CPCG01N33/577G01N33/74C07K16/26C07K2317/622C07K2317/52C07K2317/24G01N2333/58G01N2800/325
Inventor 徐淞胡川闽秦意力范舒普秀琼刘静艾海燕
Owner CHONGQING TANSHENG TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com