Journal ArticleDOI
Generation of high-affinity human antibodies by combining donor-derived and synthetic complementarity-determining-region diversity
Rene Hoet,Edward H. Cohen,Rachel Baribault Kent,Kristin L. Rookey,Sonia Schoonbroodt,Shannon Hogan,Louise Rem,Nicolas Frans,Marc Daukandt,Henk Pieters,Rob van Hegelsom,Nicole Coolen-van Neer,Nastri Horacio G,Isaac J. Rondon,Jennifer A. Leeds,Simon E. Hufton,Lili Huang,Irina Kashin,Mary Devlin,Guannan Kuang,Mieke Steukers,Malini Viswanathan,Andrew E. Nixon,Daniel J. Sexton,Hennie R. Hoogenboom,Robert Charles Ladner +25 more
TLDR
The construction of human Fab libraries having a unique combination of immunoglobulin sequences captured from human donors and synthetic diversity in key antigen contact sites in heavy-chain complementarity-determining regions 1 and 2 are reported.Abstract:
Combinatorial libraries of rearranged hypervariable V(H) and V(L) sequences from nonimmunized human donors contain antigen specificities, including anti-self reactivities, created by random pairing of V(H)s and V(L)s. Somatic hypermutation of immunoglobulin genes, however, is critical in the generation of high-affinity antibodies in vivo and occurs only after immunization. Thus, in combinatorial phage display libraries from nonimmunized donors, high-affinity antibodies are rarely found. Lengthy in vitro affinity maturation is often needed to improve antibodies from such libraries. We report the construction of human Fab libraries having a unique combination of immunoglobulin sequences captured from human donors and synthetic diversity in key antigen contact sites in heavy-chain complementarity-determining regions 1 and 2. The success of this strategy is demonstrated by identifying many monovalent Fabs against multiple therapeutic targets that show higher affinities than approved therapeutic antibodies. This very often circumvents the need for affinity maturation, accelerating discovery of antibody drug candidates.read more
Citations
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Journal ArticleDOI
Selecting and screening recombinant antibody libraries
TL;DR: The first antibody of this new generation, adalimumab (Humira, a human IgG1 specific for human tumor necrosis factor (TNF)), already approved for therapy and with many more in clinical trials, these recombinant antibody technologies will provide a solid basis for the discovery of antibody-based biopharmaceuticals, diagnostics and research reagents for decades to come.
Journal ArticleDOI
Is there new hope for therapeutic matrix metalloproteinase inhibition
TL;DR: The recent advances made in understanding the role of MMPs in inflammatory diseases and the therapeutic potential of M Parliamentary metalloproteinases inhibition in those conditions are discussed.
Journal ArticleDOI
Survey of the year 2003 commercial optical biosensor literature
Rebecca L. Rich,David G. Myszka +1 more
TL;DR: In this overview, 13 papers that should be on everyone's ‘must read’ list for 2003 are spotlighted and examples of how to identify and interpret high‐quality biosensor data are provided.
Journal ArticleDOI
Precise determination of the diversity of a combinatorial antibody library gives insight into the human immunoglobulin repertoire
Jacob Glanville,Wenwu Zhai,Jan Berka,Dilduz Telman,Gabriella Huerta,Gautam R. Mehta,Irene Ni,Li Mei,Purnima Sundar,Giles Day,David Cox,Arvind Rajpal,Jaume Pons +12 more
TL;DR: A general method for assessing human antibody sequence diversity displayed on phage using massively parallel pyrosequencing, a novel application of Kabat column-labeled profile Hidden Markov Models, and translated complementarity determining region (CDR) capture-recapture analysis is presented.
Journal ArticleDOI
Designing Human Antibodies by Phage Display
André Frenzel,Jonas Kügler,Saskia Helmsing,Doris Meier,Thomas Schirrmann,Michael Hust,Stefan Dübel +6 more
TL;DR: Some opportunities and achievements are summarized, e.g., the generation of antibodies which could not be generated otherwise, and the design of antibody properties by different panning strategies, including the adjustment of kinetic parameters.
References
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A Laboratory manual
TL;DR: To develop a program to print the barcodes using two commonly uses command sets and hence evaluates their ease of use for such applications, students should be able to program dot matrix printers, by manipulating bit level information and ink jet printers using page description language, such as PCL.
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High efficiency transformation of E.coli by high voltage electroporation
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Journal ArticleDOI
Assembly of combinatorial antibody libraries on phage surfaces: the gene III site.
TL;DR: A phagemid system was developed for the monovalent display of combinatorial antibody Fab libraries on the surface of filamentous phage M13, and may replace current antibody cloning techniques.
Journal ArticleDOI
By-passing immunization: building high affinity human antibodies by chain shuffling.
James D. Marks,Andrew D. Griffiths,Magnus Malmqvist,Tim Clackson,Jacqueline M. Bye,Greg Winter +5 more
TL;DR: Improved the affinity of one such “primary” antibody is improved by sequentially replacing the heavy and light chain variable (V) region genes with repertoires of V–genes (chain shuffling) obtained from unimmunized donors.
Journal ArticleDOI
Fully synthetic human combinatorial antibody libraries (HuCAL) based on modular consensus frameworks and CDRs randomized with trinucleotides.
Achim Knappik,Liming Ge,Annemarie Honegger,Peter Pack,Melanie Fischer,Günter Wellnhofer,Adolf Hoess,Joachim Wölle,Andreas Plückthun,Bernhard Virnekäs +9 more
TL;DR: The small number of 49 master genes will allow future improvements to be incorporated quickly, and the separation of the frameworks may help in analyzing why nature has evolved these distinct subfamilies of antibody germline genes.