# Core B: Antigen Receptor Sequencing, Cloning, Expression, and Analysis Core

> **NIH NIH U19** · UNIVERSITY OF ALABAMA AT BIRMINGHAM · 2024 · $341,791

## Abstract

Project Summary
The B cell compartment is comprised of many individual cells, each of which may express a distinct B cell
receptor (BCR). Upon antigen engagement, B cells proliferate resulting in clonal expansion, and may further
diversify their BCR genes through somatic hypermutation (SHM) and class switch recombination. These two
processes allow for affinity maturation toward the insulting antigen and the distribution of Ab reactivity across
isotypes which interface with other components of the immune system defining an Abs effector function. Clonal
expansion and diversification result in the presence of expanded groups of B cells with similar BCRs (clonal
lineages). The overarching goal of this program is to investigate the evolution and durability of immune responses
to allo, self, and foreign antigens. Although sequencing of BCR genes allows the visualization of cell expansions
and diversification that occur during the evolution of B cell responses, sequence information alone is insufficient
to evaluate changes in antigen binding that occur as immune responses evolve. The Antigen Receptor
Sequencing, Cloning, Expression, and Analysis Core (Core B) will facilitate the objectives of this program
by allowing the analysis of these two distinct but related features of humeral immunity. We will support the aims
of the projects through the following activities: i) Generating antigen receptor sequence information. Projects
require repertoire level sequence information form bulk and sorted lymphocytes, BCR for Projects 1 and 3, TCR
for Project 2. Core B will centralize the processing and quality control involved in generating next-generation of
antigen receptor libraries. We will coordinate with the Projects and with Core D, the Data and Informatics Service
Core, for analysis. ii) Generating Abs by cloning and expressing the BCR of single-sorted antigen-reactive B
cells as recombinant Ab. Both Projects 1 and 3 of this proposal are evaluating the reactivity of antigen-reactive
B cells to a variety of specificities, allo- and auto-HLA, and viral pathogen derived antigens. Using a high-
throughput Ab cloning and expression system, Core B will produce recombinant Abs from the cloned BCR of
single-sorted antigen reactive B cells across reactive with these antigens. iii) Evaluating the specificity, relative
affinity, and cross reactivity of Abs. Because understanding the effects of SHM, the relation of antigen reactivity
within and across expanded lineages, the relative affinity of Abs to their antigens and related structures requires
physical measurements of binding, Core B will evaluate Abs reactivity profiles using multiplexed cytometric
arrays. iv) Core B will generate Ab and Fab expression vectors to support the bio-physical measurements of Ab
binding and structural studies performed by Core C. Collectively, these activities will promote synergy between
the Projects and the Cores of this proposal and accelerate the research by providing access to spec...

## Key facts

- **NIH application ID:** 10824853
- **Project number:** 1U19AI181105-01
- **Recipient organization:** UNIVERSITY OF ALABAMA AT BIRMINGHAM
- **Principal Investigator:** Rodney Glenn King
- **Activity code:** U19 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $341,791
- **Award type:** 1
- **Project period:** 2024-04-16 → 2029-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10824853

## Citation

> US National Institutes of Health, RePORTER application 10824853, Core B: Antigen Receptor Sequencing, Cloning, Expression, and Analysis Core (1U19AI181105-01). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10824853. Licensed CC0.

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