A novel synthetic naïve human antibody library allows the isolation of antibodies against a new epitope of oncofetal fibronectin

Villa, Alessandra; Lovato, Valeria; Bujak, Emil; Wulhfard, Sarah; Pasche, Nadine; Neri, Dario

doi:10.4161/mabs.3.3.15616

Cited by 24 publications

(39 citation statements)

References 62 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Protein purification and size exclusion chromatography. The fusion proteins were purified by protein A affinity chromatography (HiTrap Protein A HP, GE Healthcare Life Sciences), essentially as described previously (52). Purified fusion proteins were analyzed by size exclusion chromatography on a Superdex200 10/300GL column (GE Healthcare).…”

Section: Methodsmentioning

confidence: 99%

Antibody-mediated delivery of VEGF-C potently reduces chronic skin inflammation

Schwager¹,

Renner²,

Hemmerle³

et al. 2018

JCI Insight

Self Cite

View full text Add to dashboard Cite

show abstract

Section: Methodsmentioning

confidence: 99%

Antibody-mediated delivery of VEGF-C potently reduces chronic skin inflammation

Schwager¹,

Renner²,

Hemmerle³

et al. 2018

JCI Insight

Self Cite

View full text Add to dashboard Cite

show abstract

“…Sequence variability in heavy and light chain was introduced in CDR3 loops by PCR using partially degenerated primers ( Figure S1 ; all primers were purchased from Operon), as described earlier [20], [21] and shown in Figure 1 . Briefly, DP47-based VH domains were randomly mutated from residue 95 to 100; this CDR3 loop was designed to be 4–7 amino acids long.…”

Section: Methodsmentioning

confidence: 99%

A Highly Functional Synthetic Phage Display Library Containing over 40 Billion Human Antibody Clones

et al. 2014

Self Cite

View full text Add to dashboard Cite

Several synthetic antibody phage display libraries have been created and used for the isolation of human monoclonal antibodies. The performance of antibody libraries, which is usually measured in terms of their ability to yield high-affinity binding specificities against target proteins of interest, depends both on technical aspects (such as library size and quality of cloning) and on design features (which influence the percentage of functional clones in the library and their ability to be used for practical applications). Here, we describe the design, construction and characterization of a combinatorial phage display library, comprising over 40 billion human antibody clones in single-chain fragment variable (scFv) format. The library was designed with the aim to obtain highly stable antibody clones, which can be affinity-purified on protein A supports, even when used in scFv format. The library was found to be highly functional, as >90% of randomly selected clones expressed the corresponding antibody. When selected against more than 15 antigens from various sources, the library always yielded specific and potent binders, at a higher frequency compared to previous antibody libraries. To demonstrate library performance in practical biomedical research projects, we isolated the human antibody G5, which reacts both against human and murine forms of the alternatively spliced BCD segment of tenascin-C, an extracellular matrix component frequently over-expressed in cancer and in chronic inflammation. The new library represents a useful source of binding specificities, both for academic research and for the development of antibody-based therapeutics.

show abstract

“…scFv, IgG) for selective targeting of neovascular structures in animal models (Borsi et al, 2002) and patients with cancer (Santimaria et al, 2003). In another study, 2H7 mAb, specific to a non-overlapping epitope of EDA which is distinct from previously characterized anti-EDA scFv F8, was isolated from a different version of ETH2-GOLD phage library (PHILO library), and then both scFvs (F8 and 2H7) were used to construct phage display library of chelating recombinant antibody (CRAbs) with varied inter-scFv linkers (Villa et al, 2011). It was shown that the isolated anti-EDA CRAbs (i.e.…”

Section: Pal-derived Therapeutic Mabsmentioning

confidence: 99%

Phage antibody display libraries: a powerful antibody discovery platform for immunotherapy

Zhao

Tohidkia

Siegel

et al. 2014

Critical Reviews in Biotechnology

View full text Add to dashboard Cite

Phage display technology (PDT), a combinatorial screening approach, provides a molecular diversity tool for creating libraries of peptides/proteins and discovery of new recombinant therapeutics. Expression of proteins such as monoclonal antibodies (mAbs) on the surface of filamentous phage can permit the selection of high affinity and specificity therapeutic mAbs against virtually any target antigen. Using a number of diverse selection platforms (e.g. solid phase, solution phase, whole cell and in vivo biopannings), phage antibody libraries (PALs) from the start point provides great potential for the isolation of functional mAb fragments with diagnostic and/or therapeutic purposes. Given the pivotal role of PDT in the discovery of novel therapeutic/diagnostic mAbs, in the current review, we provide an overview on PALs and discuss their impact in the advancement of engineered mAbs.

show abstract

A novel synthetic naïve human antibody library allows the isolation of antibodies against a new epitope of oncofetal fibronectin

Cited by 24 publications

References 62 publications

Antibody-mediated delivery of VEGF-C potently reduces chronic skin inflammation

Antibody-mediated delivery of VEGF-C potently reduces chronic skin inflammation

A Highly Functional Synthetic Phage Display Library Containing over 40 Billion Human Antibody Clones

Phage antibody display libraries: a powerful antibody discovery platform for immunotherapy

Contact Info

Product

Resources

About