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Amplification method of SARS-CoV-2 coronavirus antibody light chain full-length sequence

A coronavirus and antibody light chain technology, applied in the field of biomedicine, can solve the problems of weak antibody effectiveness and slow research and development progress

Inactive Publication Date: 2021-03-12
江苏中方基因生物医学科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For more than a year, the global research on antibodies has paid more and more attention. However, the research and development of antibodies in the latest published studies are all based on CDR3 fragments. This is also a major reason for the weak effectiveness of antibodies and slow progress in research and development.

Method used

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  • Amplification method of SARS-CoV-2 coronavirus antibody light chain full-length sequence

Examples

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Embodiment 1

[0018] 1. Extraction of total RNA from the blood of SARS-CoV-2 coronavirus patients or recovered patients.

[0019] Take 1.0ml of whole blood and add red blood cell lysate, centrifuge to remove the supernatant, add 1ml of lysate Trizol, and let stand at room temperature for 5 minutes. Add 200µl chloroform to shake for 15 seconds, leave at room temperature for 2 minutes, and then centrifuge at 12000rpm at 4°C for 5 minutes. Pipette the supernatant clear liquid into a clean RNase-free centrifuge tube, add an equal volume of isopropanol to mix, let stand at room temperature for 5 minutes, and centrifuge at 12,000 rpm at 4°C for 5 minutes. Discard the supernatant and add 1ml of 75% ethanol, turn it upside down for 1min, and centrifuge at 12000rpm at 4°C for 5 minutes. Discard the supernatant and dry at room temperature for 5 minutes, add 40ul RNase-free water to fully dissolve the RNA, and freeze the obtained RNA at -80°C until use.

[0020] 2. Prepare cDNA by reverse transcript...

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Abstract

The invention provides an amplification method of an SARSCoV2 coronavirus antibody light chain full-length sequence, which comprises the following steps: carrying out reverse transcription according to a 3 '-terminal constant region specific primer to obtain a full-length transcript cDNA containing a complete variable region, and carrying out PCR amplification on the cDNA prepared by reverse transcription by using the 5'-terminal variable region specific primer and a constant region primer to obtain the full-length light chain sequence. The invention provides a simple, quick and efficient method for specifically amplifying the full-length sequence of the light chain of the anti-SARSCoV2 coronavirus antibody from the blood of a patient infected with the SARSCoV2 coronavirus, and lays a foundation for deep research of the coronavirus antibody.

Description

technical field [0001] The invention relates to the field of biomedical technology, in particular to a method for amplifying the full-length light chain sequence of a SARS-CoV-2 coronavirus antibody. Background technique [0002] Coronaviruses are a class of single-stranded positive-sense RNA viruses that can infect humans and various vertebrates. They are important pathogens that cause common colds and upper respiratory tract infections in humans, and pose a serious threat to human health. The SARS-CoV-2 coronavirus that spread at the end of 2019 has the characteristics of rapid transmission, severe epidemic situation and rapid progress. Caused progressive impairment of respiratory function and death in some patients. Current drug research on SARS-CoV-2 coronavirus includes vaccines, antibodies, etc. For more than a year, the global research on antibodies has paid more and more attention. However, the research and development of antibodies in the latest published studies ...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12Q1/686C12Q2521/107
Inventor 戴方平陈东生程艳兵张韦唯
Owner 江苏中方基因生物医学科技有限公司
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