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A kind of PD-L1 antibody and its extraction method

A PD-L1, antibody technology, applied in the field of biomedicine or biopharmaceuticals, can solve the problems of narrow linear range and limited detection ability of PD-L1 antibody

Active Publication Date: 2021-09-14
深圳海创生物技术有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] However, the current traditional antibody-based PD-L1 detection has two shortcomings. First, its sensitivity is only about 50ng, and second, its linear range is relatively narrow, between 50 and 200ng. The current PD-L1 antibody detection ability is limited.

Method used

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  • A kind of PD-L1 antibody and its extraction method
  • A kind of PD-L1 antibody and its extraction method
  • A kind of PD-L1 antibody and its extraction method

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specific Embodiment approach 1

[0015] Specific embodiment 1: The nucleotide sequence of the PD-L1 antibody in this embodiment is shown in SEQ ID NO.1, and the amino acid sequence of the PD-L1 antibody is shown in SEQ ID NO.2.

specific Embodiment approach 2

[0016] Specific embodiment 2: The extraction method of PD-L1 antibody in this embodiment is carried out according to the following steps: use human PD-L1 (programmed death ligand 1) full-length protein as an antigen to immunize sharks, then extract RNA, and use reverse The cDNA was reverse-transcribed into cDNA by the recording kit, and the gene encoding the vNAR region was obtained by PCR amplification, and then the PD-L1 antibody was obtained through screening, expression and purification.

specific Embodiment approach 3

[0017] Embodiment 3: This embodiment differs from Embodiment 2 in that the upstream primer for PCR library construction is: TCGCTACCGTGGCCCAGGCGGCCAACTTGAACAAACGGGCACC, and the downstream primer for library construction is:

[0018] TGATGGTGCTGGCCGGCCTGGCCTTCATGGGTCAGAATCATGC. Other steps and parameters are the same as in the second embodiment.

[0019] Specific embodiment three: the difference between this embodiment and specific embodiment two is the PCR amplification reaction conditions: 95°C for 2 minutes; 94°C for 30 seconds, 58°C for 30 seconds, 72°C for 1 minute, 35 cycles, 72°C for 10 minutes . Other steps and parameters are the same as in the second embodiment.

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Abstract

A PD‑L1 antibody and its extraction method relate to an antibody and its extraction method. The nucleotide sequence of the PD-L1 antibody of the present invention is shown in SEQ ID NO.1, and the amino acid sequence of the PD-L1 antibody is shown in SEQ ID NO.2; the extraction method of the present invention is: using human PD-L1 (programmed death Ligand 1) The full-length protein was used as the antigen to immunize sharks, and then RNA was extracted and reverse-transcribed into cDNA using a reverse transcription kit. The gene encoding the vNAR region was amplified by PCR, and then PD‑ L1 antibody. The PD-L1 of the present invention has a small molecular weight, only 15KD, and a simple structure. The sensitivity of the antibody reaches 1ng, which greatly improves the detection ability, and the linear range is increased to 1 to 300ng, which greatly improves the range of adaptability to the detection ability, and the signal value is greatly improved. , from more than 500 to more than 2000, the PD-L1 antibody (shark antibody) obtained by the method of the present invention has improved the detection ability of traditional antibodies and increased the accuracy of detection.

Description

technical field [0001] The invention relates to the technical field of biomedicine or biopharmaceuticals, in particular to an antibody and an extraction method thereof. Background technique [0002] An antibody is a receptor that binds to an antigen, and refers to an immunoglobulin produced by the body under the stimulation of an antigenic substance, which can specifically bind to the corresponding antigen. The antibody structure consists of two heavy chains (H chains) and two light chains (L chains) (heavy chains and light chains are connected by disulfide bonds), and each chain can be divided into variable regions (V regions ) and the constant region (C region), in which the variable region has three regions with highly variable amino acid composition and arrangement sequence, called hypervariable region (HVR), because the hypervariable region can form a precise pattern with the antigenic determinant in the spatial structure Therefore, the hypervariable region is also cal...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/28C07K16/00
CPCC07K16/005C07K16/2827
Inventor 陈建明陈锦霖赵铁铭
Owner 深圳海创生物技术有限公司
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