Methods of sequencing the immune repertoire

A technology of immune group library and sequencing, which is applied in the fields of biochemical equipment and methods, recombinant DNA technology, microbial determination/testing, etc.

Active Publication Date: 2015-12-23
LINEAGE BIOSCI
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  • Abstract
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Problems solved by technology

[0007] Although reference is made to binding specificity, and indeed a large number of serological assays are based on the physical interaction between antigen and receptor, the underlying c

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  • Methods of sequencing the immune repertoire
  • Methods of sequencing the immune repertoire
  • Methods of sequencing the immune repertoire

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Embodiment Construction

[0022] The present invention provides improved methods for sequencing immune repertoires. Previous methods of determining immune repertoires, such as those described in WO2011 / 140433 and WO2012 / 083069, are based on multiplex PCR. Multiplex PCR has a number of limitations that make it particularly unsuitable for precisely determining the immune repertoire. (See figure 2 ) These limitations include capture bias and amplification bias due to PCR. Multiplex PCR techniques for sequencing immune repertoires use primers designed to prime all framework regions of known V gene segments. Capture bias occurs when mutations occur at priming sites, and genes with mutations are underamplified. PCR bias arises due to uneven gene amplification due to the relative amounts of each primer and PCR replicates with the same sequence. Thus, PCR bias can lead to apparent lack of clonality or diversity. In general, the observed repertoire is either inaccurate, or it is a linear representation of...

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Abstract

The invention provides a non-invasive technique for the detection and quantification of the immune repertoire, in a biological sample containing a plurality of distinct cell populations. Methods are conducted using sequencing technology to detect and enumerate immune repertoire within a heterogeneous biological sample.

Description

[0001] related application [0002] This application claims the benefit and priority of U.S. Provisional Application No. 61 / 806143, filed March 28, 2013, and U.S. Provisional Application No. 61 / 801785, filed March 15, 2013, the entire contents of which are incorporated by reference This article. technical field [0003] The present invention relates to the field of quantitative nucleic acid analysis. More specifically, the present invention provides methods for determining immune repertoires using high throughput sequencing. Background technique [0004] The hallmark of the adaptive immune response is the ability to generate a broad diversity of binding molecules, such as T cell antigen receptors and antibodies. Various molecular mechanisms can be used to generate initial diversity, including genetic recombination at multiple loci. Naive B cells and naive T cells are equipped with an initial repertoire of binding moieties that circulate at positions where they can come in...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12Q1/68
CPCC12N15/1096B01J19/0046B01J2219/00596B01J2219/00722C12Q2525/155C12Q2563/179C12Q2521/101C12Q2525/173C12Q2521/107C12Q1/6869C12Q2525/179C12Q1/6874C12Q1/6881C12Q2600/16
Inventor 爱德华·A·哈钦斯海-姆·克里斯蒂娜·范
Owner LINEAGE BIOSCI
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