Beyond natural antibodies: the power of in vitro display technologies

In vitro display technologies, best exemplified by phage and yeast display, were first described for the selection of antibodies some 20 years ago. Since then, many antibodies have been selected and improved upon using these methods. Although it is not widely recognized, many of the antibodies derived using in vitro display methods have properties that would be extremely difficult, if not impossible, to obtain by immunizing animals. The first antibodies derived using in vitro display methods are now in the clinic, with many more waiting in the wings. Unlike immunization, in vitro display permits the use of defined selection conditions and provides immediate availability of the sequence encoding the antibody. The amenability of in vitro display to high-throughput applications broadens the prospects for their wider use in basic and applied research.

[1]  N. Hynes,et al.  Intracellular expression of single chain antibodies reverts ErbB-2 transformation. , 1994, The Journal of biological chemistry.

[2]  J. Scott,et al.  Searching for peptide ligands with an epitope library. , 1990, Science.

[3]  T. Gerngross,et al.  Glycosylation engineering in yeast: the advent of fully humanized yeast. , 2007, Current opinion in biotechnology.

[4]  H. Steven Wiley,et al.  Flow-cytometric isolation of human antibodies from a nonimmune Saccharomyces cerevisiae surface display library , 2003, Nature Biotechnology.

[5]  C. Eigenbrot,et al.  High-affinity human antibodies from phage-displayed synthetic Fab libraries with a single framework scaffold. , 2004, Journal of molecular biology.

[6]  U. Nielsen,et al.  Selection of tumor-specific internalizing human antibodies from phage libraries. , 2000, Journal of molecular biology.

[7]  D. Burton,et al.  Detection of antibodies against the four subtypes of ebola virus in sera from any species using a novel antibody-phage indicator assay. , 2002, Virology.

[8]  P. Jackson,et al.  Disruption of Anthrax Toxin Binding with the Use of Human Antibodies and Competitive Inhibitors , 1999, Infection and Immunity.

[9]  Y. Guan,et al.  Heterosubtypic Neutralizing Monoclonal Antibodies Cross-Protective against H5N1 and H1N1 Recovered from Human IgM+ Memory B Cells , 2008, PloS one.

[10]  Boguslaw Stec,et al.  Structural and functional bases for broad-spectrum neutralization of avian and human influenza A viruses , 2009, Nature Structural &Molecular Biology.

[11]  Andreas Plückthun,et al.  Intracellular Kinase Inhibitors Selected from Combinatorial Libraries of Designed Ankyrin Repeat Proteins* , 2005, Journal of Biological Chemistry.

[12]  H. Fuchs,et al.  Targeted tumor therapies at a glance. , 2009, Current drug targets.

[13]  M. Hafner,et al.  Functional knockdown of VCAM-1 at the posttranslational level with ER retained antibodies. , 2009, Journal of immunological methods.

[14]  L. Presta,et al.  'Knobs-into-holes' engineering of antibody CH3 domains for heavy chain heterodimerization. , 1996, Protein engineering.

[15]  K. Takkinen,et al.  Fine tuning of an anti-testosterone antibody binding site by stepwise optimisation of the CDRs. , 1998, Immunotechnology : an international journal of immunological engineering.

[16]  S. Spicer,et al.  Some ascites monoclonal antibody preparations contain contaminants that bind to selected Golgi zones or mast cells. , 1994, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[17]  C. Hardman,et al.  Modelling the human immune response: performance of a 1011 human antibody repertoire against a broad panel of therapeutically relevant antigens. , 2008, Protein engineering, design & selection : PEDS.

[18]  Eugen Damoc,et al.  Detection and Purification of Tyrosine-sulfated Proteins Using a Novel Anti-sulfotyrosine Monoclonal Antibody* , 2006, Journal of Biological Chemistry.

[19]  P. S. Andersen,et al.  Shared fine specificity between T-cell receptors and an antibody recognizing a peptide/major histocompatibility class I complex. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[20]  D. Neri,et al.  Antibody-based vascular tumor targeting. , 2010, Recent results in cancer research. Fortschritte der Krebsforschung. Progres dans les recherches sur le cancer.

[21]  N. Zou,et al.  Human Single-Chain Fv Antibodies Against Burkholderia mallei and Burkholderia pseudomallei , 2007, Experimental biology and medicine.

[22]  Andreas Plückthun,et al.  Picomolar affinity antibodies from a fully synthetic naive library selected and evolved by ribosome display , 2000, Nature Biotechnology.

[23]  Sachdev S Sidhu,et al.  Synthetic antibodies from a four-amino-acid code: a dominant role for tyrosine in antigen recognition. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[24]  D. Deperthes,et al.  Streptabody, a high avidity molecule made by tetramerization of in vivo biotinylated, phage display-selected scFv fragments on streptavidin. , 2000, Molecular immunology.

[25]  R. Chiaraluce,et al.  A semi-synthetic repertoire of intrinsically stable antibody fragments derived from a single-framework scaffold. , 2001, Journal of molecular biology.

[26]  A. Murzin,et al.  The 2.0-A resolution crystal structure of a trimeric antibody fragment with noncognate VH-VL domain pairs shows a rearrangement of VH CDR3. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[27]  James A J Fitzpatrick,et al.  Fluorogen-activating single-chain antibodies for imaging cell surface proteins , 2008, Nature Biotechnology.

[28]  Matthias Paschke,et al.  Phage display systems and their applications , 2006, Applied Microbiology and Biotechnology.

[29]  L. Fugger,et al.  Recombinant antibodies with the antigen-specific, MHC restricted specificity of T cells: novel reagents for basic and clinical investigations and immunotherapy. , 1999, Immunotechnology : an international journal of immunological engineering.

[30]  Camellia W. Adams,et al.  An efficient route to human bispecific IgG , 1998, Nature Biotechnology.

[31]  M. Feldmann,et al.  Therapeutic antibodies elicited by immunization against TNF-alpha. , 1999, Nature biotechnology.

[32]  I. Kumagai,et al.  Human Anti-gold Antibodies , 2008, Journal of Biological Chemistry.

[33]  S. Dübel,et al.  A human scFv antibody generation pipeline for proteome research. , 2011, Journal of biotechnology.

[34]  J. Aster,et al.  Modulation of Notch Signaling by Antibodies Specific for the Extracellular Negative Regulatory Region of NOTCH3* , 2008, Journal of Biological Chemistry.

[35]  M. Little,et al.  A surface expression vector for antibody screening. , 1991, Gene.

[36]  J. Osbourn,et al.  From rodent reagents to human therapeutics using antibody guided selection. , 2005, Methods.

[37]  M. Foley,et al.  Green fluorescent antibodies: novel in vitro tools. , 2000, Protein engineering.

[38]  F. Gueneau,et al.  Specificity tuning of antibody fragments to neutralize two human chemokines with a single agent , 2009, mAbs.

[39]  M. Peipp,et al.  Effector mechanisms of therapeutic antibodies against ErbB receptors. , 2008, Current opinion in immunology.

[40]  S. Dübel,et al.  Development of human antibody fragments using antibody phage display for the detection and diagnosis of Venezuelan equine encephalitis virus (VEEV) , 2008, BMC biotechnology.

[41]  Eric T. Boder,et al.  Yeast surface display for screening combinatorial polypeptide libraries , 1997, Nature Biotechnology.

[42]  T. W. Smith,et al.  Kinetics in interactions between antibodies and haptens. , 1975, Biochemistry.

[43]  D. Exley,et al.  Antisera highly specific for 17 -oestradiol. , 1971, Steroids.

[44]  R. Lerner,et al.  Cloning of the immunological repertoire in Escherichia coli for generation of monoclonal catalytic antibodies: construction of a heavy chain variable region-specific cDNA library. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[45]  Clifford B Saper,et al.  An open letter to our readers on the use of antibodies , 2005, The Journal of comparative neurology.

[46]  George Georgiou,et al.  Isolation and expression of recombinant antibody fragments to the biological warfare pathogen Brucella melitensis. , 2003, Journal of immunological methods.

[47]  A. Villa,et al.  A high‐affinity human monoclonal antibody specific to the alternatively spliced EDA domain of fibronectin efficiently targets tumor neo‐vasculature in vivo , 2008, International journal of cancer.

[48]  A. Koide,et al.  The fibronectin type III domain as a scaffold for novel binding proteins. , 1998, Journal of molecular biology.

[49]  U. Nielsen,et al.  Tumor targeting using anti-her2 immunoliposomes. , 2001, Journal of controlled release : official journal of the Controlled Release Society.

[50]  Franklin Peale,et al.  Variants of the Antibody Herceptin That Interact with HER2 and VEGF at the Antigen Binding Site , 2009, Science.

[51]  Avital Lev,et al.  Recombinant antibodies with MHC‐restricted, peptide‐specific, T‐cell receptor‐like specificity: new tools to study antigen presentation and TCR–peptide–MHC interactions , 2003, Journal of molecular recognition : JMR.

[52]  H R Hoogenboom,et al.  By-passing immunization. Human antibodies from V-gene libraries displayed on phage. , 1991, Journal of molecular biology.

[53]  U. Storz.  IP Issues in the Therapeutic Antibody Industry , 2010 .

[54]  E. Unanue,et al.  Characterization and quantitation of peptide-MHC complexes produced from hen egg lysozyme using a monoclonal antibody. , 1997, Immunity.

[55]  A. Kossiakoff,et al.  An engineered substance P variant for receptor-mediated delivery of synthetic antibodies into tumor cells , 2009, Proceedings of the National Academy of Sciences.

[56]  R. Stevens,et al.  Molecular evolution of antibody cross-reactivity for two subtypes of type A botulinum neurotoxin , 2007, Nature Biotechnology.

[57]  S. Sidhu,et al.  Two-state selection of conformation-specific antibodies , 2009, Proceedings of the National Academy of Sciences.

[58]  C. Benz,et al.  Anti-CD166 single chain antibody-mediated intracellular delivery of liposomal drugs to prostate cancer cells , 2007, Molecular Cancer Therapeutics.

[59]  Toshiaki Maruyama,et al.  Human antibodies from immunized donors are protective against anthrax toxin in vivo , 2003, Nature Biotechnology.

[60]  M. Guzmán,et al.  Phage-displayed antibody fragments recognizing dengue 3 and dengue 4 viruses as tools for viral serotyping in sera from infected individuals , 2009, Archives of Virology.

[61]  G. Adams,et al.  Isolation of picomolar affinity anti-c-erbB-2 single-chain Fv by molecular evolution of the complementarity determining regions in the center of the antibody binding site. , 1996, Journal of molecular biology.

[62]  Leonard A. Smith,et al.  Neutralizing human monoclonal antibodies binding multiple serotypes of botulinum neurotoxin. , 2011, Protein engineering, design & selection : PEDS.

[63]  T. Rabbitts,et al.  Selection of antibodies for intracellular function using a two-hybrid in vivo system. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[64]  M. Hensler,et al.  Stepwise in vitro affinity maturation of Vitaxin, an αvβ3-specific humanized mAb , 1998 .

[65]  T. Rabbitts,et al.  De novo production of diverse intracellular antibody libraries. , 2003, Nucleic acids research.

[66]  Jane Wilton,et al.  The remarkable flexibility of the human antibody repertoire; isolation of over one thousand different antibodies to a single protein, BLyS. , 2003, Journal of molecular biology.

[67]  M. Braunagel,et al.  Identification of scFv antibody fragments that specifically recognise the heroin metabolite 6-monoacetylmorphine but not morphine. , 2003, Journal of immunological methods.

[68]  J. Arends,et al.  Isolation of human antibodies to tumor-associated endothelial cell markers by in vitro human endothelial cell selection with phage display libraries. , 2004, Journal of immunological methods.

[69]  M. Sheng,et al.  Antibodies in haystacks: how selection strategy influences the outcome of selection from molecular diversity libraries. , 2001, Journal of immunological methods.

[70]  J. Marks,et al.  Recombinant full-length human IgG1s targeting hormone-refractory prostate cancer , 2007, Journal of Molecular Medicine.

[71]  P. Pavlík,et al.  High-throughput screening of single-chain antibodies using multiplexed flow cytometry. , 2007, Journal of proteome research.

[72]  M. Taussig,et al.  Generating recombinant antibodies to the complete human proteome. , 2010, Trends in biotechnology.

[73]  Maurizio Molinari,et al.  β-site specific intrabodies to decrease and prevent generation of Alzheimer's Aβ peptide , 2005, The Journal of cell biology.

[74]  Leonard A. Smith,et al.  Potent neutralization of botulinum neurotoxin by recombinant oligoclonal antibody , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[75]  L. Torrance,et al.  pSKAP/S: An expression vector for the production of single-chain Fv alkaline phosphatase fusion proteins. , 1999, Protein expression and purification.

[76]  George Georgiou,et al.  Passive Protection against Anthrax by Using a High-Affinity Antitoxin Antibody Fragment Lacking an Fc Region , 2005, Infection and Immunity.

[77]  Leonard A. Smith,et al.  Extraction of BoNT/A, /B, /E, and /F with a Single, High Affinity Monoclonal Antibody for Detection of Botulinum Neurotoxin by Endopep-MS , 2010, PloS one.

[78]  E. Ooi,et al.  Neutralizing human monoclonal antibody against H5N1 influenza HA selected from a Fab-phage display library , 2008, Virology Journal.

[79]  Andreas Plückthun,et al.  An antibody library for stabilizing and crystallizing membrane proteins – selecting binders to the citrate carrier CitS , 2004, FEBS letters.

[80]  B. Jensen,et al.  Ten commercial antibodies for alpha-1-adrenergic receptor subtypes are nonspecific , 2009, Naunyn-Schmiedeberg's Archives of Pharmacology.

[81]  E. S. Graham,et al.  Specific detection of CB1 receptors; cannabinoid CB1 receptor antibodies are not all created equal! , 2008, Journal of Neuroscience Methods.

[82]  F. Bikker,et al.  Detection of Sulfur Mustard Adducts in Human Callus by Phage Antibodies , 2007, Chemical biology & drug design.

[83]  Thijs J. Hagenbeek,et al.  Therapeutic antibody targeting of individual Notch receptors , 2010, Nature.

[84]  K D Wittrup,et al.  Directed evolution of antibody fragments with monovalent femtomolar antigen-binding affinity. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[85]  Tateishi Kayoko,et al.  A novel immunization procedure for production of anti-testosterone and anti-5α-dihydrotestosterone antisera of low cross-reactivity , 1980 .

[86]  A. Skerra.  Alternative non-antibody scaffolds for molecular recognition. , 2007, Current opinion in biotechnology.

[87]  G. Winter,et al.  Cloning immunoglobulin variable domains for expression by the polymerase chain reaction. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[88]  Frederic A. Fellouse,et al.  Crystal structure of full-length KcsA in its closed conformation , 2009, Proceedings of the National Academy of Sciences.

[89]  A. Cattaneo,et al.  Intracellular immunization: antibody targeting to subcellular compartments. , 1995, Trends in cell biology.

[90]  A. Gurney,et al.  Direct demonstration of MuSK involvement in acetylcholine receptor clustering through identification of agonist ScFv , 1997, Nature Biotechnology.

[91]  Bin Zhou,et al.  Human antibodies against spores of the genus Bacillus: A model study for detection of and protection against anthrax and the bioterrorist threat , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[92]  R. Siegel,et al.  Molecular evolution of antibody affinity for sensitive detection of botulinum neurotoxin type A. , 2005, Journal of molecular biology.

[93]  J. D. White,et al.  Phase I trial of recombinant immunotoxin anti-Tac(Fv)-PE38 (LMB-2) in patients with hematologic malignancies. , 2000, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[94]  P. Chames,et al.  Improving the affinity and the fine specificity of an anti-cortisol antibody by parsimonious mutagenesis and phage display. , 1998, Journal of immunology.

[95]  T. Waldmann,et al.  Phenotypic knockout of the high-affinity human interleukin 2 receptor by intracellular single-chain antibodies against the alpha subunit of the receptor. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[96]  M. Gething,et al.  Construction of intergeneric hybrids using bacteriophage P1CM: transfer of the Klebsiella aerogenes ribitol dehydrogenase gene to Escherichia coli , 1976, Journal of bacteriology.

[97]  G. Winter,et al.  Phage antibodies: filamentous phage displaying antibody variable domains , 1990, Nature.

[98]  A. Barberis,et al.  Antigen‐independent selection of stable intracellular single‐chain antibodies , 2001, FEBS letters.

[99]  A. Cattaneo,et al.  An integrated vector system for the eukaryotic expression of antibodies or their fragments after selection from phage display libraries. , 1997, Gene.

[100]  Frederic A. Fellouse,et al.  High-throughput generation of synthetic antibodies from highly functional minimalist phage-displayed libraries. , 2007, Journal of molecular biology.

[101]  K. Weber,et al.  Coiling of intermediate filaments induced by microinjection of a vimentin-specific antibody does not interfere with locomotion and mitosis. , 1981, European journal of cell biology.

[102]  M. Little,et al.  Antibodies to steroids from a small human naive IgM library , 1997, FEBS letters.

[103]  L E Williams,et al.  Minibody: A novel engineered anti-carcinoembryonic antigen antibody fragment (single-chain Fv-CH3) which exhibits rapid, high-level targeting of xenografts. , 1996, Cancer research.

[104]  V. Roschke,et al.  Generation and characterization of LymphoStat-B, a human monoclonal antibody that antagonizes the bioactivities of B lymphocyte stimulator. , 2003, Arthritis and rheumatism.

[105]  Leonard G Presta,et al.  Molecular engineering and design of therapeutic antibodies. , 2008, Current opinion in immunology.

[106]  D. Rimm,et al.  Antibody validation by quantitative analysis of protein expression using expression of Met in breast cancer as a model , 2007, Laboratory Investigation.

[107]  Annemarie Honegger,et al.  Direct in Vivo Screening of Intrabody Libraries Constructed on a Highly Stable Single-chain Framework* , 2002, The Journal of Biological Chemistry.

[108]  M. Little,et al.  Bifunctional and multimeric complexes of streptavidin fused to single chain antibodies (scFv). , 1995, Journal of immunological methods.

[109]  F. Peale,et al.  Cross-species Vascular Endothelial Growth Factor (VEGF)-blocking Antibodies Completely Inhibit the Growth of Human Tumor Xenografts and Measure the Contribution of Stromal VEGF* , 2006, Journal of Biological Chemistry.

[110]  Frederic A. Fellouse,et al.  Synthetic antibodies for specific recognition and crystallization of structured RNA , 2008, Proceedings of the National Academy of Sciences.

[111]  T. Wurch,et al.  Trends in glycosylation, glycoanalysis and glycoengineering of therapeutic antibodies and Fc-fusion proteins. , 2008, Current pharmaceutical biotechnology.

[112]  A Roberts,et al.  In vitro selection of a high affinity antibody to oestradiol using a phage display human antibody library. , 1996, Immunotechnology : an international journal of immunological engineering.

[113]  Seung Hyun Kang,et al.  Monoclonal antibodies isolated without screening by analyzing the variable-gene repertoire of plasma cells , 2010, Nature Biotechnology.

[114]  T. Pawson,et al.  Generating a panel of highly specific antibodies to 20 human SH2 domains by phage display , 2010, Protein engineering, design & selection : PEDS.

[115]  K Dane Wittrup,et al.  Isolating and engineering human antibodies using yeast surface display , 2006, Nature Protocols.

[116]  Andreas Plückthun,et al.  Ribosome display: selecting and evolving proteins in vitro that specifically bind to a target , 2007, Nature Methods.

[117]  G. S. Hunt,et al.  Mechanistic studies of the immunochemical termination of self-tolerance with unnatural amino acids , 2009, Proceedings of the National Academy of Sciences.

[118]  P. M. Deckert.  Current constructs and targets in clinical development for antibody-based cancer therapy. , 2009, Current drug targets.

[119]  A. Kortt,et al.  High avidity scFv multimers; diabodies and triabodies. , 1999, Journal of immunological methods.

[120]  J. Xiang.  Targeting cytokines to tumors to induce active antitumor immune responses by recombinant fusion proteins. , 1999, Human antibodies.

[121]  K. Janda,et al.  Selection of human antibodies against cell surface-associated oligomeric anthrax protective antigen. , 2007, Molecular immunology.

[122]  Amos Bairoch,et al.  The Sulfinator: predicting tyrosine sulfation sites in protein sequences , 2002, Bioinform..

[123]  M. Hensler,et al.  Stepwise in vitro affinity maturation of Vitaxin, an alphav beta3-specific humanized mAb. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[124]  Wolfgang Kummer,et al.  Suitability of muscarinic acetylcholine receptor antibodies for immunohistochemistry evaluated on tissue sections of receptor gene-deficient mice , 2009, Naunyn-Schmiedeberg's Archives of Pharmacology.

[125]  D R Burton,et al.  Generation of a large combinatorial library of the immunoglobulin repertoire in phage lambda. , 1989, Science.

[126]  Jan Engberg,et al.  Visualization of Myelin Basic Protein (Mbp) T Cell Epitopes in Multiple Sclerosis Lesions Using a Monoclonal Antibody Specific for the Human Histocompatibility Leukocyte Antigen (Hla)-Dr2–Mbp 85–99 Complex , 2000, The Journal of experimental medicine.

[127]  Y. Kan,et al.  Antibodies to human fetal erythroid cells from a nonimmune phage antibody library , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[128]  J. C. Love,et al.  A microengraving method for rapid selection of single cells producing antigen-specific antibodies , 2006, Nature Biotechnology.

[129]  Valsamo Anagnostou,et al.  Antibody validation. , 2010, BioTechniques.

[130]  A. Plückthun,et al.  Assembly of a functional immunoglobulin Fv fragment in Escherichia coli. , 1988, Science.

[131]  J. Rehfeld,et al.  Microscale characterization of the binding specificity and affinity of a monoclonal antisulfotyrosyl IgG antibody , 2008, Electrophoresis.

[132]  R. Wagner,et al.  Neutralization of tetanus toxin by distinct monoclonal antibodies binding to multiple epitopes on the toxin molecule , 1984, Infection and immunity.

[133]  Y. Hooks,et al.  Application of phage display to high throughput antibody generation and characterization , 2007, Genome Biology.

[134]  J. Marks,et al.  Selection of cell binding and internalizing epidermal growth factor receptor antibodies from a phage display library. , 2001, Journal of immunological methods.

[135]  T. Logtenberg,et al.  Leucine Zipper Dimerized Bivalent and Bispecific scFv Antibodies from a Semi-synthetic Antibody Phage Display Library (*) , 1996, The Journal of Biological Chemistry.

[136]  R. Kontermann.  Intrabodies as therapeutic agents. , 2004, Methods.

[137]  A. Pini,et al.  Design and Use of a Phage Display Library , 1998, The Journal of Biological Chemistry.

[138]  G. Abraham,et al.  The evolution of titer and specificity of aldosterone binding antibodies in hyperimmunized sheep. , 1972, Steroids.

[139]  A. Cattaneo,et al.  Intracellular Immunization with Cytosolic Recombinant Antibodies , 1994, Bio/Technology.

[140]  Andreas Plückthun,et al.  Allosteric inhibition of aminoglycoside phosphotransferase by a designed ankyrin repeat protein. , 2005, Structure.

[141]  P. Nilsson,et al.  Towards proteome scale antibody selections using phage display. , 2010, New biotechnology.

[142]  J. Marks.  Deciphering antibody properties that lead to potent botulinum neurotoxin neutralization , 2004, Movement disorders : official journal of the Movement Disorder Society.

[143]  R. Williams,et al.  Crystal structure of a diabody, a bivalent antibody fragment. , 1994, Structure.

[144]  N. Bec,et al.  A focused antibody library for selecting scFvs expressed at high levels in the cytoplasm , 2007, BMC biotechnology.

[145]  Jianping Yin,et al.  Synthetic anti-BR3 antibodies that mimic BAFF binding and target both human and murine B cells. , 2006, Blood.

[146]  D. Neri,et al.  Antibody-based targeting of the tumor vasculature. , 2007, Biochimica et biophysica acta.

[147]  Hennie R. Hoogenboom,et al.  Guiding the Selection of Human Antibodies from Phage Display Repertoires to a Single Epitope of an Antigen , 1994, Bio/Technology.

[148]  A. Kortt,et al.  scFv multimers of the anti-neuraminidase antibody NC10: length of the linker between VH and VL domains dictates precisely the transition between diabodies and triabodies. , 1999, Protein engineering.

[149]  M. Westphal,et al.  A Novel One-Armed Anti-c-Met Antibody Inhibits Glioblastoma Growth In vivo , 2006, Clinical Cancer Research.

[150]  J. McCafferty,et al.  Directing phage selections towards specific epitopes. , 1996, Protein engineering.

[151]  Qun Li,et al.  Generation, Characterization and Epitope Mapping of Two Neutralizing and Protective Human Recombinant Antibodies against Influenza A H5N1 Viruses , 2009, PloS one.

[152]  H. Runnels,et al.  Human monoclonal antibodies to the insulin-like growth factor 1 receptor inhibit receptor activation and tumor growth in preclinical studies , 2010, Advances in therapy.

[153]  J. Foote,et al.  Breaking the affinity ceiling for antibodies and T cell receptors. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[154]  Andreas Plückthun,et al.  Engineered proteins as specific binding reagents. , 2005, Current opinion in biotechnology.

[155]  K. Comess,et al.  A sulfated peptide segment at the amino terminus of PSGL-1 is critical for P-selectin binding , 1995, Cell.

[156]  Ulrik B Nielsen,et al.  Therapeutic efficacy of anti-ErbB2 immunoliposomes targeted by a phage antibody selected for cellular endocytosis. , 2002, Biochimica et biophysica acta.

[157]  M. Neuberger,et al.  Affinity dependence of the B cell response to antigen: a threshold, a ceiling, and the importance of off-rate. , 1998, Immunity.

[158]  S. Dübel,et al.  Multimerization domains for antibody phage display and antibody production. , 2009, New biotechnology.

[159]  A. Bradbury,et al.  Antibodies from phage antibody libraries. , 2004, Journal of immunological methods.

[160]  Hiroyuki Kishi,et al.  A rapid and efficient single-cell manipulation method for screening antigen-specific antibody–secreting cells from human peripheral blood , 2009, Nature Medicine.

[161]  R. Minter,et al.  Human monomeric antibody fragments to TRAIL-R1 and TRAIL-R2 that display potent in vitro agonism , 2009, mAbs.

[162]  K. Fry,et al.  Rapid cloning of rearranged immunoglobulin genes from human hybridoma cells using mixed primers and the polymerase chain reaction. , 1989, Biochemical and biophysical research communications.

[163]  Timo Pulli,et al.  Automated Panning and Screening Procedure on Microplates for Antibody Generation from Phage Display Libraries , 2009, Journal of biomolecular screening.

[164]  B. Goud,et al.  Recombinant Antibodies to the Small GTPase Rab6 as Conformation Sensors , 2003, Science.

[165]  E. Fenyö,et al.  Identification of a strongly activating human anti-CD40 antibody that suppresses HIV type 1 infection. , 2008, AIDS research and human retroviruses.

[166]  G. P. Smith,et al.  Filamentous fusion phage: novel expression vectors that display cloned antigens on the virion surface. , 1985, Science.

[167]  J. Hallborn,et al.  Automated screening procedure for high-throughput generation of antibody fragments. , 2002, BioTechniques.

[168]  Bruce Tidor,et al.  Computational design of antibody-affinity improvement beyond in vivo maturation , 2007, Nature Biotechnology.

[169]  A. Plückthun,et al.  Engineering novel binding proteins from nonimmunoglobulin domains , 2005, Nature Biotechnology.

[170]  K. Peter,et al.  Subtractive single-chain antibody (scFv) phage-display: tailoring phage-display for high specificity against function-specific conformations of cell membrane molecules , 2007, Nature Protocols.

[171]  C. Bertozzi,et al.  Using Phage Display to Select Antibodies Recognizing Post-translational Modifications Independently of Sequence Context* , 2006, Molecular & Cellular Proteomics.

[172]  J. Marks,et al.  Internalizing cancer antibodies from phage libraries selected on tumor cells and yeast-displayed tumor antigens. , 2010, Journal of molecular biology.

[173]  M. Little,et al.  Di‐, tri‐ and tetrameric single chain Fv antibody fragments against human CD19: effect of valency on cell binding , 1999, FEBS letters.

[174]  Y. Iba,et al.  Monoclonal antibodies in man that neutralized H3N2 influenza viruses were classified into three groups with distinct strain specificity: 1968-1973, 1977-1993 and 1997-2003. , 2010, Virology.

[175]  D R Burton,et al.  CDR walking mutagenesis for the affinity maturation of a potent human anti-HIV-1 antibody into the picomolar range. , 1995, Journal of molecular biology.

[176]  Jennifer S. Martinez,et al.  Selection and characterization of scFv antibodies against the Sin Nombre hantavirus nucleocapsid protein. , 2007, Journal of immunological methods.

[177]  D. Collins,et al.  Comparative specificity of antisera raised against estrone, estradiol-17 and estriol using 6-0-carboxy-methyloxime bovine serum albumin derivatives. , 1973, Steroids.

[178]  George Georgiou,et al.  Protection against anthrax toxin by recombinant antibody fragments correlates with antigen affinity , 2002, Nature Biotechnology.