Scholarlink

<p>The human antibody repertoire is a promising source for therapeutic-grade antibodies. Yet current methods for strategically mining these B cell repertoires are stymied by throughput and chain pairing considerations. This study presents advancements in fluidics and molecular biology that enable the multi-step encapsulation and capture of B cells from an immunized, humanized mouse in nanoliter sized droplets. Once singularly captured, antigen-specific B-cells can be lysed and individually manipulated via RT-PCR to splice cognate V genes and create a predominantly natively paired library. To explore the importance of these process improvements in library generation, we constructed natively-paired libraries against two therapeutically-relevant human proteins. Through deep sequencing, bioinformatics-driven screening and phage display, we selected functional, target-specific antibodies. Our findings reveal that natively paired libraries contain a higher percentage of target-specific antibodies and demonstrate enhanced potency and improved developability in both in silico and in vitro assessments relative to combinatorial library-derived antibodies. Furthermore, antibodies with native pairing show increased potency as well as improved in silico and in vitro developability compared to their randomly paired counterparts. To this end, we see this droplet microfluidic platform and its capacity to generate and facilitate the high-throughput interrogation of antigen-specific antibody repertoires as an important, orthogonal therapeutic antibody discovery approach.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

A droplet microfluidics-based platform for generating target-specific, natively-paired immune libraries and identifying potent and developable antibodies

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年1月3日

查看全文
  • Anusuya Ramasubramanian,
  • Brendan T. Deveney,
  • Sarah A. Clark,
  • Paula Boutin,
  • Kayla Keepseagle,
  • Saleh Riahi,
  • Taotao Lao,
  • Megan Salemi,
  • Sagar Kathuria,
  • Pep Amengual-Rigo,
  • Dilyana Dimova,
  • Leighton Marcovici,
  • Joern Hopke,
  • Jocelyn Newton,
  • Kalie Mix,
  • Kevin Kramer,
  • Hélène Erasimus,
  • Norbert Furtmann,
  • Guillaume Mottet,
  • Nikolai Suslov,
  • Ankit Mahendra,
  • Sambasiva Rao Pemmaraju,
  • Partha S. Chowdhury,
  • David A. Weitz,
  • John A. Heyman

摘要

The human antibody repertoire is a promising source for therapeutic-grade antibodies. Yet current methods for strategically mining these B cell repertoires are stymied by throughput and chain pairing considerations. This study presents advancements in fluidics and molecular biology that enable the multi-step encapsulation and capture of B cells from an immunized, humanized mouse in nanoliter sized droplets. Once singularly captured, antigen-specific B-cells can be lysed and individually manipulated via RT-PCR to splice cognate V genes and create a predominantly natively paired library. To explore the importance of these process improvements in library generation, we constructed natively-paired libraries against two therapeutically-relevant human proteins. Through deep sequencing, bioinformatics-driven screening and phage display, we selected functional, target-specific antibodies. Our findings reveal that natively paired libraries contain a higher percentage of target-specific antibodies and demonstrate enhanced potency and improved developability in both in silico and in vitro assessments relative to combinatorial library-derived antibodies. Furthermore, antibodies with native pairing show increased potency as well as improved in silico and in vitro developability compared to their randomly paired counterparts. To this end, we see this droplet microfluidic platform and its capacity to generate and facilitate the high-throughput interrogation of antigen-specific antibody repertoires as an important, orthogonal therapeutic antibody discovery approach.