# US 12,180,304 B2 · Variable region sequence of broad-spectrum antibody against clothianidin and dinotefuran and preparation of intact recombinant antibody thereof

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- Number: US 12,180,304 B2
- Document ID: US12180304B2
- Status: Granted
- Kind: B2
- Application number: 17/443,888
- Filed: 2021-07-28
- Published: 2022-02-03
- Granted: 2024-12-31
- Earliest claimed: 2020-07-28
- Assignees: Zhejiang University
- Applicants: Zhejiang University
- Inventors: Yirong Guo; Yunyun Chang; Ying Zhao; Pengyan Liu; Ying Liu; Yang Chen; Guonian Zhu
- Web page: https://patentgrep.com/patent/US12180304B2

## CPC

- C07K 16/44: against material not provided for elsewhere {, e.g. haptens, metals, DNA, RNA, amino acids}
- C07K 2317/20: characterized by taxonomic origin
- C07K 2317/33: Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
- C07K 2317/64: comprising a combination of variable region and constant region components
- C07K 2317/92: Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)

## Abstract

The present disclosure provides a variable region sequence of a broad-spectrum antibody against clothianidin and dinotefuran, where a gene encoding a heavy chain variable region has an amino acid sequence shown in SEQ ID NO: 2. The present disclosure further discloses a broad-spectrum intact recombinant antibody against clothianidin and dinotefuran, including a heavy chain constant region, a heavy chain variable region, a light chain constant region, and a light chain variable region, where a gene encoding the heavy chain variable region has an amino acid sequence shown in SEQ ID NO: 2. The sequence genes obtained by the present disclosure are ligated to an expression vector containing a heavy chain constant region gene and a light chain constant region gene, respectively, and an intact recombinant antibody is expressed and obtained by using mammalian cells with a double-plasmid system.

## Claims

### Claim 1

A broad-spectrum intact recombinant antibody against clothianidin and dinotefuran, comprising a heavy chain constant region, a heavy chain variable region comprising an amino acid sequence of SEQ ID NO:2, a light chain constant region, and a light chain variable region comprising an amino acid sequence of SEQ ID NO: 4.

### Claim 2 (depends on 1)

The broad-spectrum intact recombinant antibody against clothianidin and dinotefuran according to claim 1, wherein a gene encoding the heavy chain variable region has a nucleotide sequence shown in SEQ ID NO: 1.

### Claim 3 (depends on 1)

The broad-spectrum intact recombinant antibody against clothianidin and dinotefuran according to claim 1, wherein a gene encoding the light chain variable region has a nucleotide sequence shown in SEQ ID NO: 3.

### Claim 4 (depends on 1)

The broad-spectrum intact recombinant antibody against clothianidin and dinotefuran according to claim 1, wherein a gene encoding the heavy chain variable region has a nucleotide sequence shown in SEQ ID NO: 1 and a gene encoding the light chain variable region has a nucleotide sequence shown in SEQ ID NO: 3.

### Claim 5 (depends on 1)

An expression plasmid for expressing the broad-spectrum intact recombinant antibody against clothianidin and dinotefuran of claim 1.

### Claim 6 (depends on 5)

The expression plasmid of claim 5, wherein a gene encoding the heavy chain variable region has a nucleotide sequence shown in SEQ ID NO: 1.

### Claim 7 (depends on 5)

The expression plasmid of claim 5, wherein a gene encoding the light chain variable region has a nucleotide sequence shown in SEQ ID NO:3.

### Claim 8 (depends on 5)

The expression plasmid of claim 5, wherein a gene encoding the heavy chain variable region has a nucleotide sequence shown in SEQ ID NO: 1 and a gene encoding the light chain variable region has a nucleotide sequence shown in SEQ ID NO: 3.

### Claim 9 (depends on 5)

The expression plasmid of claim 5, comprising a mouse IgG1 heavy chain constant region nucleotide sequence, the heavy chain variable region nucleotide sequence shown in SEQ ID NO: 1, a mouse kappa light chain constant region nucleotide sequence and a light chain variable region nucleotide sequence shown in SEQ ID NO: 3, wherein the nucleotide sequence of the heavy chain constant region and the heavy chain variable region is capable of expressing the heavy chain protein of the broad-spectrum intact recombinant antibody against clothianidin and dinotefuran, and the nucleotide sequence of the light chain constant region and the light chain variable region is capable of expressing the light chain protein of the broad-spectrum intact recombinant antibody against clothianidin and dinotefuran.