Special Issue "Antibody Engineering"

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A special issue of Antibodies (ISSN 2073-4468).

Deadline for manuscript submissions: closed (20 September 2014)

Special Issue Editor

Guest Editor
Dr. Thomas Böldicke (Website)

Recombinant Protein Expression/Intrabody Unit, Helmholtz-Centre for Infection Research, Inhoffenstraße 7, D 38124 Braunschweig, Germany
Interests: antibody engineering; selection of human antibodies; cancer therapy; intracellular antibodies

Special Issue Information

Dear Colleagues,

Antibody engineering has revolutionized the current generation of antibodies, which are needed for detection of proteins, analysis and inhibition of protein functions, as well as diagnostic and therapeutic purposes. Human, chimeric and humanized antibodies with high specificity and affinity can now be generated by antibody engineering. Chimeric or humanized variants of mouse antibodies are applied in the clinic. Fusion of antibodies with cytokines, immunotoxins, specific Fc receptors and different signal peptides can be engineered. Phage display of large collections of antibody fragments enables the selection of specific antibodies in vitro. Many human antibodies selected by phage display are being studied and are showing promise in clinical trials.

This Special Issue on antibody engineering will present new technology developments and in vitro and in vivo applications of new engineered antibodies. Researchers will present their latest results and Review articles will cover new developments in the field.

Dr. Thomas Böldicke
Guest Editors

Submission

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. Papers will be published continuously (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are refereed through a peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Antibodies is an international peer-reviewed Open Access quarterly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 300 CHF (Swiss Francs). English correction and/or formatting fees of 250 CHF (Swiss Francs) will be charged in certain cases for those articles accepted for publication that require extensive additional formatting and/or English corrections.


Keywords

  • human antibody libraries
  • cell surface display systems
  • ribosomal and mRNA display
  • expression and purification of recombinant antibodies
  • bispecific antibodies
  • immunotoxins
  • immunocytokines
  • intracellular antibodies
  • therapeutic antibodies
  • in vitro affinity maturation
  • improving folding and stability of recombinant antibodies

Published Papers (4 papers)

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Research

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Open AccessArticle Codon-Precise, Synthetic, Antibody Fragment Libraries Built Using Automated Hexamer Codon Additions and Validated through Next Generation Sequencing
Antibodies 2015, 4(2), 88-102; doi:10.3390/antib4020088
Received: 5 March 2015 / Revised: 21 April 2015 / Accepted: 11 May 2015 / Published: 15 May 2015
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Abstract
We have previously described ProxiMAX, a technology that enables the fabrication of precise, combinatorial gene libraries via codon-by-codon saturation mutagenesis. ProxiMAX was originally performed using manual, enzymatic transfer of codons via blunt-end ligation. Here we present Colibra™: an automated, proprietary version of [...] Read more.
We have previously described ProxiMAX, a technology that enables the fabrication of precise, combinatorial gene libraries via codon-by-codon saturation mutagenesis. ProxiMAX was originally performed using manual, enzymatic transfer of codons via blunt-end ligation. Here we present Colibra™: an automated, proprietary version of ProxiMAX used specifically for antibody library generation, in which double-codon hexamers are transferred during the saturation cycling process. The reduction in process complexity, resulting library quality and an unprecedented saturation of up to 24 contiguous codons are described. Utility of the method is demonstrated via fabrication of complementarity determining regions (CDR) in antibody fragment libraries and next generation sequencing (NGS) analysis of their quality and diversity. Full article
(This article belongs to the Special Issue Antibody Engineering)
Open AccessArticle Reduced Culture Temperature Differentially Affects Expression and Biophysical Properties of Monoclonal Antibody Variants
Antibodies 2014, 3(3), 253-271; doi:10.3390/antib3030253
Received: 24 July 2014 / Revised: 15 August 2014 / Accepted: 21 August 2014 / Published: 29 August 2014
Cited by 1 | PDF Full-text (1130 KB) | HTML Full-text | XML Full-text
Abstract
Reduced culture temperature is an increasingly popular practice to improve recombinant protein yields in CHO cells. Recent studies have attributed the enhancement of protein titers at sub-physiological temperatures to increased mRNA levels as well as extended stationary phase. We observed that reducing [...] Read more.
Reduced culture temperature is an increasingly popular practice to improve recombinant protein yields in CHO cells. Recent studies have attributed the enhancement of protein titers at sub-physiological temperatures to increased mRNA levels as well as extended stationary phase. We observed that reducing the culture temperature arrested cell growth, prolonged viability, and increased cell size. However, the reduced culture temperature had a differential effect on protein and mRNA expression of closely related antibody mutants from stable cell lines. The highly expressing mutant (Ala) exhibited similar or decreased specific productivity and decreased volumetric productivity over the culture lifetime at 32 °C compared to 37 °C. In contrast, the specific and volumetric productivity of the poorly expressing mutant (Gly) was enhanced at the lower culture temperature. The difference in specific productivity was reflected in the amounts of heavy- and light-chain mRNA. Analysis of the secondary and tertiary configurations of the purified antibodies by circular dichroism revealed fundamental structural differences imposed by the Ala to Gly mutation as well as reduced culture temperature. We propose that the effect of reduced culture temperature on expression is protein-dependent; protein folding fidelity and assembly is improved at lower temperatures, enhancing the expression of proteins that have a propensity to misfold. Full article
(This article belongs to the Special Issue Antibody Engineering)
Open AccessArticle Kinetic Characterization of a Panel of High-Affinity Monoclonal Antibodies Targeting Ricin and Recombinant Re-Formatting for Biosensor Applications
Antibodies 2014, 3(2), 215-231; doi:10.3390/antib3020215
Received: 19 February 2014 / Revised: 10 April 2014 / Accepted: 28 April 2014 / Published: 9 May 2014
Cited by 1 | PDF Full-text (1099 KB) | HTML Full-text | XML Full-text
Abstract
Ricin is a potent glycoprotein toxin that is structurally composed of two subunits joined via a disulfide bond: a ~30 kDa subunit A (RTA) and a ~32 kDa subunit B (RTB). There are fears of ricin being used as a weapon for [...] Read more.
Ricin is a potent glycoprotein toxin that is structurally composed of two subunits joined via a disulfide bond: a ~30 kDa subunit A (RTA) and a ~32 kDa subunit B (RTB). There are fears of ricin being used as a weapon for warfare and terrorism and, as such, there is an increasing need for the development of immunodiagnostic reagents targeted towards this toxin. This article describes the production and characterization of a panel of six ricin-specific monoclonal IgG antibodies (mAbs), previously selected based upon their ability to inhibit ricin-mediated killing of cultured cells. Subsequent epitope binding analysis using the surface plasmon resonance (SPR) array biosensor (ProteOn XPR36) indicated three distinct, non-competitive binding epitopes (“bins”). The association (ka) and dissociation (kd) rate constants and binding affinities (KD) of each of the mAbs to ricin were also determined by SPR using Biacore T100 instrument. Affinities (KD) ranged from 0.1 nM to 9 nM. We present the coding sequences of the variable domains of the six mAbs, the expression, kinetic and cytotoxicity assays for two recombinant Fab (rFab) fragments and demonstrate a rFab affinity improvement by chain-shuffling. Together, these antibodies and constituent rFabs represent a panel of reagents for high-affinity recognition of ricin with potential national security biosensor applications. Full article
(This article belongs to the Special Issue Antibody Engineering)
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Review

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Open AccessReview Engineered Bovine Antibodies in the Development of Novel Therapeutics, Immunomodulators and Vaccines
Antibodies 2014, 3(2), 205-214; doi:10.3390/antib3020205
Received: 24 February 2014 / Revised: 5 April 2014 / Accepted: 25 April 2014 / Published: 9 May 2014
Cited by 3 | PDF Full-text (428 KB) | HTML Full-text | XML Full-text
Abstract
Some bovine antibodies across all classes are unique, such as the CDR3 of the variable heavy-domain (VH CDR3), which is exceptionally long (up to 66 amino acids), unlike most conventional antibodies where the VH CDR3 loops range from 10 to 25 amino [...] Read more.
Some bovine antibodies across all classes are unique, such as the CDR3 of the variable heavy-domain (VH CDR3), which is exceptionally long (up to 66 amino acids), unlike most conventional antibodies where the VH CDR3 loops range from 10 to 25 amino acids. The exceptionally long VH CDR3 is encoded by unusually long germline IGHD genes together with insertion of novel “a” nucleotide rich conserved short nucleotide sequence (CSNS) specifically at the IGH V-D junction. Such an exceptionally long VH CDR3 confers unique “knob and stalk” structural architecture where the knob, formed by intra-VH CDR3 disulfide bridges, is separated by 20 Å solvent exposed stalk composed of anti-parallel beta strands. The substitution of the knob with cytokines, such as, erythropoietin and granulocyte colony stimulating factor 3 (granulocyte colony stimulating factor), results in expression of functional fusion proteins with enhanced pharmacokinetics. The beta stranded stalk can be substituted with other rigid structures, for example, repeat alpha helices to form coiled-coil that mimics the beta-stranded stalk and, thus, opens opportunities for insertion of this structure in the CDRs of antibodies across species. Given the versatility of such a structural platform in bovine antibody VH CDR3, it provides the opportunity for the development of new generation of diagnostics, therapeutics, vaccines and immunomodulating drugs. Full article
(This article belongs to the Special Issue Antibody Engineering)
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