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https://www.readbyqxmd.com/read/29227825/engineering-of-single-chain-antibodies-for-solubility
#1
Yaghoub Safdari
In the current study, we introduce a method to design intrinsically soluble single chain antibodies (scFvs) that can be easily produced in bacterial expression systems in a soluble form. Summarily, CDR loops are grafted on framework regions derived from intrinsically soluble subclass 3 (VH3 and VL3) human germline sequences. Framework-donor sequences should carry CDR loops of interest (in terms of canonical classes) and contain special residues in their hydrophobic cores. Recombinant variable fragments resultant from CDR grafting are subjected to 3D modeling, mutated (if necessary), and superposed to parental variable domains...
December 8, 2017: International Immunopharmacology
https://www.readbyqxmd.com/read/29219059/inhibition-of-intercellular-communication-between-prostate-cancer-cells-by-a-specific-anti-steap-1-single-chain-antibody
#2
Seyed-Alireza Esmaeili, Foroogh Nejatollahi, Amirhossein Sahebkar
BACKGROUND: Six-Transmembrane epithelial antigen of the prostate-1 (STEAP-1) is present at the intercellular junctions of the secretory epithelium of prostate and is overexpressed in all steps of prostate cancer. STEAP-1 acts as a transporter protein or a putative channel between cancer cells while it has limited expression in normal human tissues. This protein has been suggested as an attractive target for prostate cancer immunotherapy. OBJECTIVE: This study aimed at the development of a specific single chain fragment variable (scFv) antibody against STEAP-1 epitope and testing the inhibitory effect of the selected scFv antibody in blocking gap junctions between tumor cells...
December 7, 2017: Anti-cancer Agents in Medicinal Chemistry
https://www.readbyqxmd.com/read/29213270/nanobodies-and-nanobody-based-human-heavy-chain-antibodies-as-antitumor-therapeutics
#3
REVIEW
Peter Bannas, Julia Hambach, Friedrich Koch-Nolte
Monoclonal antibodies have revolutionized cancer therapy. However, delivery to tumor cells in vivo is hampered by the large size (150 kDa) of conventional antibodies. The minimal target recognition module of a conventional antibody is composed of two non-covalently associated variable domains (VH and VL). The proper orientation of these domains is mediated by their hydrophobic interface and is stabilized by their linkage to disulfide-linked constant domains (CH1 and CL). VH and VL domains can be fused via a genetic linker into a single-chain variable fragment (scFv)...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/29200285/site-specific-modification-of-single-chain-antibody-fragments-for-bioconjugation-and-vascular-immunotargeting
#4
Colin Fred Greineder, Carlos H Villa, Landis R Walsh, Raisa Y Kiseleva, Elizabeth D Hood, Makan Khoshnejad, Robert Warden-Rothman, Andrew Tsourkas, Vladimir R Muzykantov
Conjugation of antibodies to drugs and drug carriers improves delivery to target tissues. Widespread implementation and effective translation of this pharmacologic strategy awaits development of affinity ligands capable of a defined degree of modification and highly efficient bioconjugation without loss of affinity. To date, such ligands are lacking for the targeting of therapeutics to vascular endothelial cells. To enable site-specific click chemistry conjugation to therapeutic cargo, we used the bacterial transpeptidase, sortase A, to attach short azidolysine containing peptides to three endothelial-specific single chain antibody fragments (scFv)...
December 4, 2017: Bioconjugate Chemistry
https://www.readbyqxmd.com/read/29199989/crystal-structure-of-an-anti-idiotype-variable-lymphocyte-receptor
#5
Bernard C Collins, Hiro Nakahara, Sharmistha Acharya, Max D Cooper, Brantley R Herrin, Ian A Wilson
Variable lymphocyte receptors (VLRs), the leucine-rich repeat (LRR)-based antigen receptors of jawless fish, have great utility in a wide variety of biochemical and biological applications, similar to classical Ig-based antibodies. VLR-based reagents may be particularly useful when traditional antibodies are not available. An anti-idiotype lamprey VLR, VLR39, has previously been identified that recognizes the heavy-chain CDR3 of the B-cell receptor (BCR) of a leukemic clone from a patient with chronic lymphocytic leukemia (CLL)...
December 1, 2017: Acta Crystallographica. Section F, Structural Biology Communications
https://www.readbyqxmd.com/read/29199242/a-single-step-breeding-generated-a-diagnostic-anti-cortisol-antibody-fragment-with-over-30-fold-enhanced-affinity
#6
Hiroyuki Oyama, Izumi Morita, Yuki Kiguchi, Tomomi Morishita, Sakiko Fukushima, Yuki Nishimori, Toshifumi Niwa, Norihiro Kobayashi
Cortisol levels in bodily fluids represent a useful index for pituitary-adrenal function, and thus practical anti-cortisol antibodies are required. We have studied "antibody-breeding" approaches, which involve in vitro evolution of antibodies to improve their antigen-binding performances. Here, we produced an antibody fragment to measure serum cortisol levels with over 30-fold enhanced affinity after single mutagenesis and selection steps. A mouse anti-cortisol antibody, Ab-CS#3, with insufficient affinity for practical use, was chosen as the prototype antibody...
2017: Biological & Pharmaceutical Bulletin
https://www.readbyqxmd.com/read/29189116/scfv-improvement-approaches
#7
Mozafar Mohammadi, Peyman Bemani, Ali Hakakian
The most common recombinant antibody format is the single chain fragment variable (scFv) which it contains the complete antigen-binding domains of an intact antibody. ScFv fragments have found vast medical and non-medical applications. Several approaches have been employed to increase the affinity, avidity and structural stability related to these antibody fragments. Most approaches related to scFv improvement have been included in this review.
November 29, 2017: Protein and Peptide Letters
https://www.readbyqxmd.com/read/29188305/synergistic-cytotoxicity-of-a-prostate-cancer-specific-immunotoxin-in-combination-with-the-bh3-mimetic-abt-737
#8
Theresa Noll, Susanne Schultze-Seemann, Irina Kuckuck, Marta Michalska, Philipp Wolf
In many tumors, including prostate cancer, anti-apoptotic members of the Bcl-2 family are overexpressed and cause cell death resistance, which is a typical hallmark of cancer. Different therapeutic approaches, therefore, aim to restore the death mechanisms for enhanced apoptosis. Our recombinant immunotoxin D7(VL-VH)-PE40 is composed of the scFv D7(VL-VH) against the prostate-specific membrane antigen (PSMA) on the surface of prostate cancer cells and of the cytotoxic domain of the bacterial toxin Pseudomonas Exotoxin A (PE40)...
November 29, 2017: Cancer Immunology, Immunotherapy: CII
https://www.readbyqxmd.com/read/29187855/cd4-binding-site-directed-cross-neutralizing-scfv-monoclonals-from-hiv-1-subtype-c-infected-indian-children
#9
Sanjeev Kumar, Rajesh Kumar, Lubina Khan, Muzamil Ashraf Makhdoomi, Ramachandran Thiruvengadam, Madhav Mohata, Mudit Agarwal, Rakesh Lodha, Sushil Kumar Kabra, Subrata Sinha, Kalpana Luthra
Progression of human immunodeficiency virus type-1 (HIV-1) infection in children is faster than adults. HIV-1 subtype C is responsible for more than 50% of the infections globally and more than 90% infections in India. To date, there is no effective vaccine against HIV-1. Recent animal studies and human Phase I trials showed promising results of the protective effect of anti-HIV-1 broadly neutralizing antibodies (bnAbs). Interaction between CD4 binding site (CD4bs) on the HIV-1 envelope glycoprotein and CD4 receptor on the host immune cells is the primary event leading to HIV-1 infection...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/29186655/engineering-bifunctional-antibodies-with-constant-region-fusion-architectures
#10
Juanjuan Du, Yu Cao, Yan Liu, Ying Wang, Yong Zhang, Guangsen Fu, Yuhan Zhang, Lucy Lu, Xiaozhou Luo, Chan Hyuk Kim, Peter G Schultz, Feng Wang
We report a method to generate bifunctional antibodies by grafting full-length proteins into constant region loops of a full-length antibody or an antigen-binding fragment (Fab). The fusion proteins retain the antigen binding activity of the parent antibody but have an additional activity associated with the protein insert. The engineered antibodies have excellent in-vitro activity, physiochemical properties, and stability. Among these, a Her2×CD3 bispecific antibody (BsAb) was constructed by inserting an anti-Her2 ScFv into an anti-CD3 Fab...
November 29, 2017: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/29185855/production-of-a-germline-humanized-cetuximab-scfv-and-evaluation-of-its-activity-in-recognizing-egfr-overexpressing-cancer-cells
#11
Arsham Banisadr, Yaghoub Safdari, Anvarsadat Kianmehr, Mahdieh Pourafshar
The aim of this study was to produce a humanized single chain antibody (scFv) as a potential improved product design to target EGFR (Epidermal Growth Factor Receptor) overexpressing cancer cells. To this end, CDR loops of cetuximab (an FDA-approved anti-EGFR antibody)were grafted on framework regions derived from type 3 (VH3 and VL3 kappa)human germline sequences to obtain recombinant VH and VL domainslinked together with a flexible linker [(Gly4Ser)3]to form a single chain antibody (scFv). Codon optimized synthetic gene encoding the scFv (with NH2-VH-linker-VL-COOH orientation) was expressed in E...
November 29, 2017: Human Vaccines & Immunotherapeutics
https://www.readbyqxmd.com/read/29183788/reduction-of-nonspecificity-motifs-in-synthetic-antibody-libraries
#12
Ryan L Kelly, Doris Le, Jessie Zhao, K Dane Wittrup
Successful antibody development requires both functional binding and desirable biophysical characteristics. In the current study we analyze the causes of one hurdle to clinical development, off-target reactivity or nonspecificity. We used a high-throughput nonspecificity assay to isolate panels of nonspecific antibodies from two synthetic scFv libraries expressed on the surface of yeast, identifying both individual amino acids and motifs within the complementarity determining regions (CDRs) which contribute to the phenotype...
November 25, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/29180536/high-affinity-gd2-specific-car-t-cells-induce-fatal-encephalitis-in-a-preclinical-neuroblastoma-model
#13
Sarah A Richman, Selene Nunez-Cruz, Babak Moghimi, Lucy Z Li, Zachary T Gershenson, Zissimos Mourelatos, David M Barrett, Stephan A Grupp, Michael C Milone
The GD2 ganglioside, which is abundant on the surface of neuroblastoma cells, is targeted by an FDA-approved therapeutic monoclonal antibody and is an attractive tumor-associated antigen for cellular immunotherapy. Chimeric antigen receptor (CAR)-modified T cells can have potent antitumor activity in B-cell malignancies, and trials to harness this cytolytic activity toward GD2 in neuroblastoma are underway. In an effort to enhance the antitumor activity of CAR T cells that target GD2, we generated variant CAR constructs predicted to improve the stability and the affinity of the GD2-binding, 14G2a-based, single-chain variable fragment (scFv) of the CAR, and compared their properties in vivo...
November 27, 2017: Cancer Immunology Research
https://www.readbyqxmd.com/read/29178403/n-glycan-engineering-of-a-plant-produced-anti-cd20-hil-2-immunocytokine-significantly-enhances-its-effector-functions
#14
Carla Marusic, Claudio Pioli, Szymon Stelter, Flavia Novelli, Chiara Lonoce, Elena Morrocchi, Eugenio Benvenuto, Anna Maria Salzano, Andrea Scaloni, Marcello Donini
Anti-CD20 recombinant antibodies are among the most promising therapeutics for the treatment of B-cell malignancies such as non-Hodgkin lymphomas. We recently demonstrated that an immunocytokine (2B8-Fc-hIL2), obtained by fusing an anti-CD20 scFv-Fc antibody derived from C2B8 mAb (rituximab) to the human interleukin 2 (hIL-2), can be efficiently produced in Nicotiana benthamiana plants. The purified immunocytokine (IC) bearing a typical plant protein N-glycosylation profile showed a CD20 binding activity comparable to that of rituximab and was efficient in eliciting antibody-dependent cell-mediated cytotoxicity (ADCC) of human PBMC against Daudi cells, indicating its fuctional integrity...
November 27, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29167497/development-and-validation-of-scfv-conjugated-affinity-silk-protein-for-specific-detection-of-carcinoembryonic-antigen
#15
Mitsuru Sato, Hiroshi Kitani, Katsura Kojima
The production costs for monoclonal antibodies (MAbs) utilized in medical diagnostic kits are inevitably high because the MAbs are mostly obtained from hybridoma cell culture. Here, we report the development and validation of a novel affinity silk protein produced by transgenic silkworm technology as a possible alternative diagnostic tool for cancers. We generated a transgenic silkworm expressing a cDNA construct containing fibroin L-chain fused to a single-chain variable fragment (scFv) derived from a MAb against carcinoembryonic antigen (CEA)...
November 22, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29165857/isolation-of-phage-display-library-derived-scfv-antibody-specific-to-listeria-monocytogenes-by-a-novel-immobilized-method
#16
Nguyen Xuan-Hung, Thu-Le Trinh, Huong Thi-Bich Vu, Huan Quang Le, Anh To Kim
AIMS: To select Listeria monocytogenes specific scFv antibodies from a phage display library by a novel simple and cost-effective immobilization method. METHODS AND RESULTS: Light Expanded Clay Aggregate (LECA) was used as biomass support matrix for biopanning of a phage display library to select Listeria monocytogenes specific scFv antibody. Four rounds of positive selection against LECA-immobilized L. monocytogenes and an additional subtractive panning against L...
November 22, 2017: Journal of Applied Microbiology
https://www.readbyqxmd.com/read/29165352/antibody-based-protective-immunity-against-helminth-infections-antibody-phage-display-derived-antibodies-against-bmr1-antigen
#17
Anizah Rahumatullah, Izzati Zahidah Abdul Karim, Rahmah Noordin, Theam Soon Lim
Helminth parasite infections are significantly impacting global health, with more than two billion infections worldwide with a high morbidity rate. The complex life cycle of the nematodes has made host immune response studies against these parasites extremely difficult. In this study, we utilized two phage antibody libraries; the immune and naïve library were used to identify single chain fragment variable (scFv) clones against a specific filarial antigen (BmR1). The V-gene analysis of isolated scFv clones will help shed light on preferential VDJ gene segment usage against the filarial BmR1 antigen in healthy and infected states...
November 22, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29163493/alpha-galactosylceramide-cd1d-antibody-fusion-proteins-redirect-invariant-natural-killer-t-cell-immunity-to-solid-tumors-and-promote-prolonged-therapeutic-responses
#18
REVIEW
Lianjun Zhang, Alena Donda
Major progress in cancer immunotherapies have been obtained by the use of tumor targeting strategies, in particular with the development of bi-functional fusion proteins such as ImmTacs or BiTes, which engage effector T cells for targeted elimination of tumor cells. Given the significance of invariant natural killer T (iNKT) cells in bridging innate and adaptive immunity, we have developed a bi-functional protein composed of the extracellular part of CD1d molecule that was genetically fused to an scFv fragment from high affinity antibodies against HER2 or CEA...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/29159981/understanding-the-metabolic-burden-of-recombinant-antibody-production-in-saccharomyces-cerevisiae-using-a-quantitative-metabolomics-approach
#19
Jorg C de Ruijter, Essi V Koskela, Jatin Nandania, Alexander D Frey, Vidya Velagapudi
The cellular changes induced by heterologous protein expression in the yeast Saccharomyces cerevisiae have been analyzed on many levels and found to be significant. However, even though high-level protein production poses a metabolic burden, evaluation of the expression host at the level of the metabolome has often been neglected. We present a comparison of metabolite profiles of a wild-type strain with those of three strains producing recombinant antibody variants of increasing size and complexity: a scFv fragment, a scFv-Fc fusion protein, and a full-length IgG molecule...
November 20, 2017: Yeast
https://www.readbyqxmd.com/read/29159220/data-on-atherosclerosis-specific-antibody-conjugation-to-nanoemulsions
#20
Geoffrey Prévot, Martine Duonor-Cérutti, Mélusine Larivière, Jeanny Laroche-Traineau, Marie Josée Jacobin-Valat, Philippe Barthélémy, Gisèle Clofent-Sanchez, Sylvie Crauste-Manciet
This article present data related to the publication entitled "Iron oxide core oil-in-water nanoemulsion as tracer for atherosclerosis MPI and MRI imaging" (Prévot et al., 2017) [1]. Herein we describe the engineering in the baculovirus-insect cell system and purification processes of the human scFv-Fc TEG4-2C antibody, specific of platelets within the atheroma plaque. For molecular targeting purpose, atheroma specific antibody was conjugated to nanoemulsions (NEs) using a heterobifunctional linker (DSPE-PEG-maleimide)...
December 2017: Data in Brief
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