A Two-Step Golden Gate Cloning Procedure for the Generation of Natively Paired YSD Fab Libraries.
Bidirectional promoter
Fab library
Golden Gate cloning
Paired VH–VL antibody library
Restriction enzyme type IIs
Yeast surface display
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2023
2023
Historique:
medline:
7
7
2023
pubmed:
5
7
2023
entrez:
5
7
2023
Statut:
ppublish
Résumé
In vitro antibody display libraries have emerged as powerful tools for a streamlined discovery of novel antibody binders. While in vivo antibody repertoires are matured and selected as a specific pair of variable heavy and light chains (VH and VL) with optimal specificity and affinity, during the recombinant generation of in vitro libraries, the native sequence pairing is not maintained. Here we describe a cloning method that combines the flexibility and versatility of in vitro antibody display with the advantages of natively paired VH-VL antibodies. In this regard, VH-VL amplicons are cloned via a two-step Golden Gate cloning procedure, allowing the display of Fab fragments on yeast cells.
Identifiants
pubmed: 37405648
doi: 10.1007/978-1-0716-3279-6_10
doi:
Substances chimiques
Antibodies
0
Immunoglobulin Fab Fragments
0
Peptide Library
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
161-173Informations de copyright
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
Références
Charles J, Janeway A, Travers P, Walport M, Shlomchik MJ (2001) The structure of a typical antibody molecule. In: The immune system in health and disease, 5th edn. Garland Science
Fernández-Quintero ML et al (2020) Antibodies exhibit multiple paratope states influencing VH–VL domain orientations. Commun Biol 3(1):1–14
doi: 10.1038/s42003-020-01319-z
Hoogenboom HR (2005) Selecting and screening recombinant antibody libraries. Nat Biotechnol 23(9):1105–1116
doi: 10.1038/nbt1126
pubmed: 16151404
Chen Y, Kim SH, Shang Y, Guillory J, Stinson J, Zhang Q, Hötzel I, Hoi KH (2018) Barcoded sequencing workflow for high throughput digitization of hybridoma antibody variable domain sequences. J Immunol Methods 455:88–94
doi: 10.1016/j.jim.2018.01.004
pubmed: 29357282
Tiller T, Meffre E, Yurasov S, Tsuiji M, Nussenzweig MC, Wardemann H (2008) Efficient generation of monoclonal antibodies from single human B cells by single cell RT-PCR and expression vector cloning. J Immunol Methods 329(1–2):112–124
doi: 10.1016/j.jim.2007.09.017
pubmed: 17996249
Rajan S et al (2018) Recombinant human B cell repertoires enable screening for rare, specific, and natively paired antibodies. Commun Biol 1(1):1–8
doi: 10.1038/s42003-017-0006-2
Adler AS et al (2018) A natively paired antibody library yields drug leads with higher sensitivity and specificity than a randomly paired antibody library. mAbs 10(3):431–443
doi: 10.1080/19420862.2018.1426422
pubmed: 29376776
pmcid: 5916548
Vaughan TJ et al (1996) Human antibodies with sub-nanomolar affinities isolated from a large non-immunized phage display library. Nat Biotechnol 14(3):309–314
doi: 10.1038/nbt0396-309
pubmed: 9630891
Ledsgaard L, Kilstrup M, Karatt-Vellatt A, McCafferty J, Laustsen AH (2018) Basics of antibody phage display technology. Toxins 10(6):236
doi: 10.3390/toxins10060236
pubmed: 29890762
pmcid: 6024766
Almagro JC, Pedraza-Escalona M, Arrieta HI, Pérez-Tapia SM (2019) Phage display libraries for antibody therapeutic discovery and development. Antibodies 8(3):44
doi: 10.3390/antib8030044
pubmed: 31544850
pmcid: 6784186
Colby DW, Kellogg BA, Graff CP, Yeung YA, Swers JS, Wittrup KD (2004) Engineering antibody affinity by yeast surface display. Methods Enzymol 388:348–358
doi: 10.1016/S0076-6879(04)88027-3
pubmed: 15289082
Baek D-S, Kim Y-S (2014) Construction of a large synthetic human Fab antibody library on yeast cell surface by optimized yeast mating. J Microbiol Biotechnol 24(3):408–420
doi: 10.4014/jmb.1401.01002
pubmed: 24394194
Steinwand M, Droste P, Frenzel A, Hust M, Dübel S, Schirrmann T (2014) The influence of antibody fragment format on phage display based affinity maturation of IgG. MAbs 6(1):204–218
doi: 10.4161/mabs.27227
pubmed: 24262918
Sivelle C, Sierocki R, Ferreira-Pinto K, Simon S, Maillere B, Nozach H (2018) Fab is the most efficient format to express functional antibodies by yeast surface display. MAbs 10(5):720–729
doi: 10.1080/19420862.2018.1468952
pubmed: 29708852
pmcid: 6150635
Weaver-Feldhaus JM, Lou J, Coleman JR, Siegel RW, Marks JD, Feldhaus MJ (2004) Yeast mating for combinatorial Fab library generation and surface display. FEBS Lett 564(1–2):24–34
doi: 10.1016/S0014-5793(04)00309-6
pubmed: 15094038
Engler C, Kandzia R, Marillonnet S (2008) A one pot, one step, precision cloning method with high throughput capability. PLoS One 3(11):e3647
doi: 10.1371/journal.pone.0003647
pubmed: 18985154
pmcid: 2574415
Engler C, Marillonnet S (2013) Combinatorial DNA assembly using Golden Gate cloning. In: Synthetic biology. Humana Press, Totowa, pp 141–156
doi: 10.1007/978-1-62703-625-2_12
Boder ET, Wittrup KD (1997) Yeast surface display for screening combinatorial polypeptide libraries. Nat Biotechnol 15(6):553–557
doi: 10.1038/nbt0697-553
pubmed: 9181578
Benatuil L, Perez JM, Belk J, Hsieh C-M (2010) An improved yeast transformation method for the generation of very large human antibody libraries. Protein Eng Des Sel 23(4):155–159
doi: 10.1093/protein/gzq002
pubmed: 20130105