Antibody phage display technology and its applications

Hoogenboom, Hennie R.; Bruïne, Adriaan P. de; Hufton, Simon E.; Hoet, Rene; Arends, Jan–Willem; Roovers, Rob C.

doi:10.1016/s1380-2933(98)00007-4

Cited by 460 publications

(239 citation statements)

References 142 publications

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“…In the majority of the examples cited, conventional hybridoma technology was used. An alternative approach is the phage-display technology, which involves positive selection in the procedure (Hoogenboom et al, 1998). Using this approach, our laboratory has screened a very large human naı¨ve antibody library against a variety of recombinant human HLA-A2-peptide complexes displaying T-cell epitopes specific for antigens expressed in melanoma, prostate, breast, ovary and lung cancer (Cohen et al, 2002;Denkberg et al, 2002a;Denkberg et al, 2003;Cohen et al, 2003a;Held et al, 2004;Denkberg and Reiter, 2006).…”

Section: T-cell Receptor-like Antibodiesmentioning

confidence: 99%

Novel antibodies as anticancer agents

2007

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In recent years antibodies, whether generated by traditional hybridoma technology or by recombinant DNA strategies, have evolved from Paul Ehrlich's 'magic bullets' to a modern age 'guided missile'. In the recent years of immunologic research, we are witnessing development in the fields of antigen screening and protein engineering in order to create specific anticancer remedies. The developments in the field of recombinant DNA, protein engineering and cancer biology have let us gain insight into many cancer-related mechanisms. Moreover, novel techniques have facilitated tools allowing unique distinction between malignantly transformed cells, and regular ones. This understanding has paved the way for the rational design of a new age of pharmaceuticals: monoclonal antibodies and their fragments. Antibodies can select antigens on both a specific and a high-affinity account, and further implementation of these qualities is used to target cancer cells by specifically identifying exogenous antigens of cancer cell populations. The structure of the antibody provides plasticity resonating from its functional sites. This review will screen some of the many novel antibodies and antibody-based approaches that are being currently developed for clinical applications as the new generation of anticancer agents.

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Section: T-cell Receptor-like Antibodiesmentioning

confidence: 99%

Novel antibodies as anticancer agents

2007

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“…27 Briefly, specific sets of degenerated primers were used to PCR-amplify the cDNA segments that correspond to the immunoglobulin heavy and light chain variable domains. The V H and V L PCR pools were assembled into a scFv repertoire by a PCR overlap extension reaction and subsequently cloned as an SfiI-NotI fragment into the pCANTAB6 phagemid vector.…”

Section: Construction and Screening Of Antibody Scfv Librarymentioning

confidence: 99%

Targeting an extracellular epitope of the human multidrug resistance protein 1 (MRP1) in malignant cells with a novel recombinant single chain Fv antibody

Binyamin¹,

Assaraf²,

Haus-Cohen³

et al. 2004

Intl Journal of Cancer

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Inherent and acquired multidrug resistance (MDR) is characterized by a simultaneous resistance to diverse anticancer drugs and is a major impediment towards curative chemotherapy of cancer. Hence one important goal is to develop strategies aimed at specific targeting of major anticancer drug efflux transporters of the ATP-binding cassette (ABC) superfamily including multidrug resistance protein 1 -MRP1 (ABCC1). To date, no monoclonal antibody has been isolated that can target an extracellular MRP1 epitope. Using a phage display approach, we have isolated a recombinant singlechain Fv (scFv) antibody that specifically reacts with the extracellular N-terminus of the human MRP1. Flow cytometric analysis revealed that this scFv fragment binds specifically to various viable human tumor cells that display variable Key words: multi drug resistance (MDR); scFv antibody; MRP1; phage displayCombination chemotherapy continues to play a major role in the treatment of various human malignancies. However, the efficacy of various chemotherapeutics has been limited by the frequent emergence of various anticancer drug resistance phenomena. The best-studied mechanisms of multidrug resistance (MDR) in malignant cells involve the overexpression of ATP-driven anticancer drug efflux pumps of the ABC superfamily. 1-3 These include the multidrug transporter, P-glycoprotein (Pgp), which extrudes various hydrophobic cytotoxic agents, as well as members of the multidrug resistance protein (MRP) family, currently comprising 9 exporters (MRP1 through MRP9). 3-5 MRP1 was discovered during a differential hybridization screen aimed at identifying mRNAs overexpressed in doxorubicin-resistant lung cancer cells that lack P-glycoprotein overexpression. 6 The 100 -200-fold MRP1 mRNA overexpression was a result of gene amplification. Analysis of the 1,531 amino acid sequence of MRP1 sequence identified this protein as a member of the ATP-binding cassette (ABC) superfamily of transporter proteins. 6 Importantly, transfection of the MRP1 cDNA conferred upon sensitive cells MDR to anthracyclines, Vinca alkaloids, etoposide, arsenical and antimonial oxyanions but not to cisplatinum and mitoxantrone. 7 These studies paved the way for the isolation of additional MRPs, MRP2 through MRP9, which have been shown to confer resistance to various toxic compounds that are either initially hydrophilic or that undergo intracellular conjugation to glutathione, sulfate and glucuronate. 5 Furthermore, another ABC transporter recently shown to mediate MDR in various tumor cells of epithelial origin is the breast cancer resistance protein (BCRP). 8 In order to study anticancer drug transporter proteins such as MRP1 in clinical specimens and to reveal their physiological functions, specific monoclonal antibodies (MAbs) were recently developed. 9 -11 Monoclonal antibodies against MDR proteins were used for 2 main purposes: a) detection and quantification of MRP and P-glycoprotein expression in drug-resistant cancer cells and b) reversal of MDR in cancer cells by abolishin...

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“…16 Briefly, specific sets of degenerated primers were used to PCR-amplify the cDNA segments corresponding to the immunoglobulin VH and VL domains. The VH and VL PCR pools were assembled into an scFv repertoire by a PCR overlap extension and subsequently cloned as an SfiI-NotI fragment into the pCANTAB6 phagemid vector.…”

Section: Construction and Screening Of Antibody Scfv Librarymentioning

confidence: 99%

“…Eluted phages were expanded in exponentially growing Escherichia coli TG1 cells, which were later superinfected with M13KO7 helper phage as described. 16 …”

Section: Construction and Screening Of Antibody Scfv Librarymentioning

confidence: 99%

“…[13][14][15] Moreover, using antibody phage display libraries, it is possible to isolate new recombinant scFv antibodies with the desired specificity and affinity directed toward a large array of antigens. 16 Our aim was to explore the possibility of using a small recombinant scFv antibody molecule for the modulation and reversal of the MDR phenotype in tumor cells by direct inhibition of Pgpmediated drug-efflux activity. Using an antibody phage display approach, we isolated a recombinant scFv fragment capable of disrupting Pgp-efflux activity.…”

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confidence: 99%

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Disruption of P‐glycoprotein anticancer drug efflux activity by a small recombinant single‐chain Fv antibody fragment targeted to an extracellular epitope

Haus-Cohen

Assaraf

Binyamin

et al. 2004

Intl Journal of Cancer

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Inherent and acquired MDR is characterized by simultaneous resistance to diverse anticancer drugs and continues to be a major impediment in the curative chemotherapy of cancer. The MDR1 gene product, Pgp, is an ATP-driven efflux pump, which extrudes a variety of dissimilar hydrophobic cytotoxic compounds from MDR cells. Pgp overexpression results in MDR of tumor cell lines in vitro as well as of a variety of human malignancies. Thus, one major goal is to develop strategies aimed at specifically disrupting Pgp drugefflux activity. To this end, we have developed a small recombinant antibody capable of potent reversal of MDR, by disrupting Pgp drug-efflux activity. Using a phage display approach, we isolated a small scFv recombinant antibody fragment that specifically reacts with the first extracellular loop of human Pgp. This scFv fragment binds specifically to various Pgp-overexpressing human MDR carcinoma cell lines, consequently disrupts Pgp drug-efflux function and thereby reverses the MDR phenotype. We have successfully disrupted anticancer drug-extrusion pump activity in MDR cells using a small recombinant scFv fragment. We propose that these novel small Fv-based recombinant antibody molecules may lead to the development of a new class of antibody fragment-based agents that specifically inhibit Pgp drug extrusion. Hence, these small recombinant antibody fragments may be applied in combination chemotherapy to overcome MDR in various human cancers. © 2004 Wiley-Liss, Inc. Key words: P-glycoprotein; multidrug resistance; single-chain FvCombination chemotherapy continues to play a major role in the treatment of various human malignancies. MDR, whereby tumor cells simultaneously possess intrinsic or acquired cross-resistance to diverse anticancer drugs, hampers the efficacy of the chemotherapeutic treatment of various human cancers. 1-3 Intense molecular investigations of the MDR phenomenon over the past 2 decades have resulted in the isolation and characterization of genes encoding for several plasma membrane glycoproteins associated with MDR. The best-studied mechanism of MDR in cancer cells is overexpression of an energy-dependent efflux pump, the multidrug transporter Pgp. 4 Pgp is a member of a large ABC superfamily of transport proteins. 5 Pgp-mediated MDR plays an important clinical role in the resistance of various tumor cells to chemotherapy, as judged by the high incidence of MDR1 expression in tumors of various types and the correlations between MDR1 expression and lack of response to chemotherapy in various human malignancies. 6,7 These findings prompted a major effort aimed at identifying strategies and agents capable of reversing Pgp-mediated MDR. 3 Inhibitors of the MDR phenotype in cancer cells may function in 2 main modes: they can either modify the expression (i.e., decrease it) or disrupt the drug-efflux function of the transporter proteins involved in MDR. 3 The search for "chemosensitizers" that disrupt the function of these drug exporters and thereby reverse MDR has grown in parallel with th...

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Antibody phage display technology and its applications

Cited by 460 publications

References 142 publications

Novel antibodies as anticancer agents

Novel antibodies as anticancer agents

Targeting an extracellular epitope of the human multidrug resistance protein 1 (MRP1) in malignant cells with a novel recombinant single chain Fv antibody

Disruption of P‐glycoprotein anticancer drug efflux activity by a small recombinant single‐chain Fv antibody fragment targeted to an extracellular epitope

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