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Identification of antigen-specific adaptive immune responses using Arm-PCR and high-throughput sequencing

A specific and antigenic technology, applied in the direction of immunoglobulin, microbial determination/testing, specific peptides, etc., can solve problems such as time-consuming, hybridoma genetic instability, animal pain and suffering, etc.

Inactive Publication Date: 2014-08-13
CB BIOTECHNOLOGIES INC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Hybridoma technology has historically been time-consuming and labor-intensive, and the resulting hybridomas may be genetically unstable (Chambers, R.S. (2005). High-throughput antibody production. Curr Opin Chem Biol 9:46-50.)
Additionally, if the hybridomas multiply in mice too much ascites can accumulate, potentially causing pain and distress to the animal

Method used

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  • Identification of antigen-specific adaptive immune responses using Arm-PCR and high-throughput sequencing
  • Identification of antigen-specific adaptive immune responses using Arm-PCR and high-throughput sequencing
  • Identification of antigen-specific adaptive immune responses using Arm-PCR and high-throughput sequencing

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Embodiment Construction

[0039] Hemagglutinin (HA) is an antigenic glycoprotein found on the surface of influenza viruses and used as a component of vaccines to elicit an immune response. The 2009-2010 formulation contained 30 μg / ml HA of each of the following three viruses: A / Brisbane / 59 / 2007, IVR-148 (H1N1), A / Uruguay / 716 / 2007, NYMC X-175C (H3N2) (class A / Brisbane / 10 / 2007 virus), and B / Brisbane / 60 / 2008. Administration of the 2009-2010 influenza vaccine to two healthy volunteers The volunteers reported feeling normal and well for a period of 30 days prior to vaccination. Both the 2008-2009 and 2009-2010 vaccines contained essentially the same influenza A H1N1 and H3N2 antigens but different B-strain antigens. To test our approach and provide the highest probability of matching antigen-specific antibodies to body fluid repertoire sequencing results, we selected volunteers who had previously received the 2008-2009 influenza vaccine.

[0040] Sample Preparation

[0041] Blood samples were taken at...

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Abstract

Disclosed is a method for correlating at least one amino acid sequence from an antibody isolated from human or animal blood with at least one DNA sequence corresponding to the antibody in the immunorepertoire of the human or animal. The method also provides a means for pairing heavy and light chains to produce synthesized monoclonal antibodies.

Description

[0001] priority statement [0002] This application claims the benefit of priority to earlier filed US Provisional Patent Application No. 61 / 540,454, filed September 28, 2011. As permitted by applicable laws and regulations, the contents of US Provisional Application No. 61 / 540,454 are incorporated herein by reference. field of invention [0003] The present invention relates to methods for identifying antigen-specific adaptive immune responses. Background of the invention [0004] Monoclonal antibodies (mAbs) are widely used in applications ranging from diagnostic and research reagents to therapeutic drugs. A critical step in the production of medically useful antibodies (Abs) is the initial identification of Abs specific for the desired antigen. This is often done using multiple rounds of "panning" in techniques such as hybridoma and phage display or by ELISPOT in chip-based methods such as ISAAC (Jin, A., et al. (2009), A rapid and efficient single-cell manipulation me...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/08C07K14/00G06F19/16
CPCC07K16/1018C12Q1/6874C07K16/00G01N33/56966G01N33/53C07K2317/55C07K2317/21C40B30/04
Inventor 韩建米兰达·伯恩-斯蒂尔
Owner CB BIOTECHNOLOGIES INC
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