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Generation of nanobodies from transgenic 'LamaMice' lacking an endogenous immunoglobulin repertoire.

Thomas EdenAlessa Z SchaffrathJanusz WesolowskiTobias StählerNatalie TodeNathalie RichterWaldemar SchäferJulia HambachIrm Hermans-BorgmeyerJannis WoensCamille Le GallSabrina WendlerChristian Linke-WinnebeckMartina StobbeIwona BudnickiAmelie WanneyYannic HeitzLena SchimmelpfennigLaura SchweitzerDennis ZimmerErik StahlFabienne SeyfriedAnna J GebhardtLynn DieckowKristoffer RieckenBoris FehsePeter BannasTim MagnusMartijn VerdoesCarl G FigdorKlaus F HartleppHubertus SchleerJonas FünerNicola M TomasFriedrich HaagBjörn RissiekAnna M MannStephan MenzelFriedrich Nolte
Published in: Nature communications (2024)
Due to their exceptional solubility and stability, nanobodies have emerged as powerful building blocks for research tools and therapeutics. However, their generation in llamas is cumbersome and costly. Here, by inserting an engineered llama immunoglobulin heavy chain (IgH) locus into IgH-deficient mice, we generate a transgenic mouse line, which we refer to as 'LamaMouse'. We demonstrate that LamaMice solely express llama IgH molecules without association to Igκ or λ light chains. Immunization of LamaMice with AAV8, the receptor-binding domain of the SARS-CoV-2 spike protein, IgE, IgG2c, and CLEC9A enabled us to readily select respective target-specific nanobodies using classical hybridoma and phage display technologies, single B cell screening, and direct cloning of the nanobody-repertoire into a mammalian expression vector. Our work shows that the LamaMouse represents a flexible and broadly applicable platform for a facilitated selection of target-specific nanobodies.
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