# US 2014/0155291 A1 · Method for Identifying Lineage-Related Antibodies

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 2014/0155291 A1
- Document ID: US20140155291A1
- Status: Published application
- Kind: A1
- Application number: 14/177,003
- Filed: 2014-02-10
- Published: 2014-06-05
- Earliest claimed: 2009-09-11
- Assignees: Epitomics, Inc. (c/o Abcam plc)
- Applicants: Epitomics, Inc. (c/o Abcam plc)
- Inventors: GUO-LIANG YU
- Web page: https://patentgrep.com/patent/US20140155291A1

## CPC

- G01N 33/577: involving monoclonal antibodies {binding reaction mechanisms characterised by the use of monoclonal antibodies (G01N33/5302 - G01N33/576 take precedence)}
- C07K 16/00: Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
- C07K 16/2863: {against receptors for growth factors, growth regulators}
- C12Q 1/6876: Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- G01N 33/566: using specific carrier or receptor proteins as ligand binding reagents {where possible specific carrier or receptor proteins are classified with their target compounds}
- G01N 33/6854: {Immunoglobulins}
- C07K 2317/10: characterized by their source of isolation or production
- C07K 2317/56: variable (Fv) region, i.e. VH and/or VL
- C07K 2317/76: Antagonist effect on antigen, e.g. neutralization or inhibition of binding
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- 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)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

In certain embodiments, the method may comprise: a) obtaining the antibody sequences from a population of B cells; b) grouping the antibody sequences to provide a plurality of groups of lineage-related antibodies; c) testing a single antibody from each of the groups in a bioassay and, after the first antibody has been identified, d) testing further antibodies that are in the same group as the first antibody in a second bioassay. In another embodiment, the method may comprise: a) testing a plurality of antibodies obtained from a first portion of an antibody producing organ of an animal; b) obtaining the sequence of a first identified antibody; c) obtaining from a second portion of said antibody producing organ the sequences of further antibodies that are related by lineage to said first antibody; and, c) testing the further antibodies in a second bioassay.

## Claims

### Claim at position 1

1-20. (canceled)

### Claim 21

A method for identifying an antibody that binds to an antigen, comprising:
a) obtaining: (i) antibody heavy chain variable domain sequences and (ii) antibody light chain variable domain sequences by sequencing heavy and light chain-encoding cDNAs made from a population of antibody-producing B cells of a rabbit that has been immunized by the antigen;
b) grouping the heavy chain variable domain sequences of step a) into a plurality of heavy chain groups and, independently, grouping the light chain variable domain sequences of step a) into a plurality of light chain groups, wherein:
(i) heavy chain sequences that comprise CDR3 regions that are of the same length and contain up to five amino acid substitutions relative to one another are grouped together; and
(ii) light chain sequences that comprise CDR3 regions that are of the same length and contain up to five amino acid substitutions relative to one another are grouped together;
c) selecting a heavy chain group and a light chain group from the groups produced by step b);
d) pairing a heavy chain variable domain sequence and a light chain variable domain sequence from the groups selected in c); and
e) testing an antibody comprising the heavy and light chain sequences paired in d) for binding to the antigen.

### Claim 22 (depends on 21)

The method of claim 21, wherein the heavy and light chain variable domain sequences paired in step d) are not naturally paired together in the rabbit of step a).

### Claim 23 (depends on 21)

The method of claim 21, wherein:
(i) heavy chain sequences that comprise CDR3 regions that are of the same length and contain 0, 1 or 2 amino acid substitutions relative to one another are grouped together; and
(ii) light chain sequences that comprise CDR3 regions that are of the same length and contain 0, 1 or 2 amino acid substitutions relative to one another are grouped together.

### Claim 24 (depends on 21)

The method of claim 21, wherein the grouping step (b) further comprises comparing the entire lengths of the heavy and light chain variable domain sequences to one another.

### Claim 25 (depends on 21)

The method of claim 21, wherein the obtaining step a) is done by sequencing cDNAs encoding the heavy and light chain variable domain sequences from single B cells, or cultures of the same.

### Claim 26 (depends on 21)

The method of claim 21, wherein the obtaining step a) is done by sequencing cDNAs encoding the heavy and light chain variable domain sequences made from a plurality of pools of B cells.

### Claim 27 (depends on 21)

The method of claim 21, wherein the heavy and light chain groups of selecting step c) are selected by analysis of lineage trees.

### Claim 28 (depends on 21)

The method of claim 21, wherein, prior to step a), the antibody-producing B cells are enriched by binding to the antigen.

### Claim 29 (depends on 21)

The method of claim 21, wherein, prior to step a), the antibody-producing B cells are not enriched by binding to the antigen

### Claim 30 (depends on 21)

The method of claim 21, wherein the selected heavy chain and light chain groups selected in step c) both contain at least 10 members.

### Claim 31 (depends on 21)

The method of claim 21, wherein the testing step e) comprises testing the antibody of e) in a blocking assay and/or a neutralization assay.

### Claim 32 (depends on 21)

The method of claim 21, wherein the testing step e) comprises testing the antibody of e) in an ELISA.

### Claim 33 (depends on 21)

The method of claim 21, wherein the obtaining step a) comprises obtaining at least 1,000 different antibody heavy chain variable domain sequences and at least 1,000 different antibody light chain variable domain sequences.

### Claim 34 (depends on 21)

The method of claim 21, wherein the antibody-producing B cells are cells obtained from spleen, lymph node or peripheral blood.