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scfv phage display

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https://www.readbyqxmd.com/read/29775561/isolation-and-characterization-of-a-novel-anti-salbutamol-chicken-scfv-for-human-doping-urinalysis
#1
Warren Lee, Syed A Ali, Leow Chiuan Yee, Tan Soo Choon, Leow Chiuan Herng
Anti-salbutamol antibodies remain as important tools for the detection of salbutamol abuse in athletic doping. This study evaluated the feasibility and efficiency of the chicken (Gallus gallus domesticus) as an immunization host to generate anti-salbutamol scFv antibodies by phage display. A phage display antibody library was constructed from a single chicken immunized against salbutamol-KLH conjugate. After a stringent biopanning strategy, a novel scFv clone which was inhibited by free salbutamol recorded the highest affinity...
May 15, 2018: Analytical Biochemistry
https://www.readbyqxmd.com/read/29753735/molecular-characterization-of-single-chain-antibody-variable-fragments-scfv-specific-to-pep27-from-streptococcus-pneumoniae
#2
Dongho Kim, ShinA Jang, Jihye Oh, Seungsu Han, Soobin Park, Prachetash Ghosh, Dong-Kwon Rhee, Sangho Lee
Pep27 from Streptococcus pneumoniae is reported to initiate pneumococcal autolysis, thereby constituting a major virulence factor. Although a few antisera recognizing Pep27 have been reported, no monoclonal, well-characterized antibody for Pep27 has been developed. Here we screened two single-chain antibody variable fragments (scFv) using a phage display from a large human synthetic scFv library to select clones E2 and F9. Dissociation constants (Kd ) of E2 and F9 were 1.1 μM and 0.50 μM, respectively...
May 10, 2018: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/29727718/preparation-of-single-chain-antibody-against-vp3-protein-of-duck-hepatitis-virus-type-1-by-phage-display-technology
#3
Yongjuan Wang, Pingfu Cui, Shanyuan Zhu, Ting Meng, Fuxing Hao, Guoqiang Zhu, Weiyong Zuo
To construct phage antibody library for VP3 protein of duck hepatitis virus type 1(DHAV-1) and pan the specific single-chain variable fragment antibody (scFv), total RNA was extracted from the protein VP3- immunized mice spleen., vp3 gene encoding VP3 protein was amplified from the genome of DHAV-1 by RT-PCR method for the following recombinant pET-VP3 construction, immunogenic VP3 expression and purification, and combined with SOE-PCR method to complete the assembly of scFv. The scFv gene was cloned into pCANTAB5E vector for phage antibody library construction...
May 1, 2018: Journal of Virological Methods
https://www.readbyqxmd.com/read/29714016/array-in-well-epitope-mapping-of-phage-displayed-antibodies
#4
Urpo Lamminmäki, Gaurav Batra, Petri Saviranta
Novel affinity reagents, such as single chain (scFv) antibody fragments, can be generated by isolating them from recombinant protein libraries using phage display selection. A successful selection process against a target protein can produce a number of binder candidates among which the desired binders are identified by screening and characterization of individual clones. Obtaining information on the binding properties, such as the binding epitope, already during the screening step helps to choose the most useful candidates for further development at early phase saving time and resources...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29678657/development-and-evaluation-of-an-optimal-human-single-chain-variable-fragment-derived-bcma-targeted-car-t-cell-vector
#5
Eric L Smith, Mette Staehr, Reed Masakayan, Ishan J Tatake, Terence J Purdon, Xiuyan Wang, Pei Wang, Hong Liu, Yiyang Xu, Sarah C Garrett-Thomson, Steven C Almo, Isabelle Riviere, Cheng Liu, Renier J Brentjens
B cell maturation antigen (BCMA) has recently been identified as an important multiple myeloma (MM)-specific target for chimeric antigen receptor (CAR) T cell therapy. In CAR T cell therapy targeting CD19 for lymphoma, host immune anti-murine CAR responses limited the efficacy of repeat dosing and possibly long-term persistence. This clinically relevant concern can be addressed by generating a CAR incorporating a human single-chain variable fragment (scFv). We screened a human B cell-derived scFv phage display library and identified a panel of BCMA-specific clones from which human CARs were engineered...
March 28, 2018: Molecular Therapy: the Journal of the American Society of Gene Therapy
https://www.readbyqxmd.com/read/29628793/challenging-tumour-immunological-techniques-that-help-to-track-cancer-stem-cells-in-malignant-melanomas-and-other-solid-tumours
#6
Beatrix Kotlan, Vanda Plotar, Klara Eles, Szabolcs Horvath, Timea Balatoni, Orsolya Csuka, Mihaly Újhelyi, Ákos Sávolt, Andras Szollar, Istvan Vamosi-Nagy, Laszlo Toth, Emil Farkas, Jozsef Toth, Miklos Kasler, Gabriella Liszkay
Aim of the study: The arsenal of questions and answers about the minor cancer initiating cancer stem cell (CSC) population put responsible for cancer invasiveness and metastases, has left with an unsolved puzzle. Specific aims of a complex project were partly focused on revealing new biomarkers of cancer. We designed and set up novel techniques to facilitate the detection of cancerous cells. Materials and methods: As a novel approach, we investigated B cells infiltrating breast carcinomas and melanomas (TIL-B) in terms of their tumour antigen binding potential...
March 2018: Contemporary Oncology Współczesna Onkologia
https://www.readbyqxmd.com/read/29627856/generation-and-characterization-of-a-novel-recombinant-scfv-antibody-specific-for-campylobacter-jejuni
#7
Ruramayi M Nzuma, Fuquan Liu, Irene R Grant
Campylobacter jejuni is a leading cause of foodborne illness worldwide, mainly due to consumption and handling of contaminated raw chicken. Rapid detection methods for C. jejuni are vital for monitoring contamination levels in chicken products and reducing human Campylobacteriosis cases. The 'gold standard' culture-based method of Campylobacter detection takes 3-5 days and is too slow to permit effective intervention. Immuno-based methods are faster, but usually necessitate use of animals or hybridoma technology to produce antibodies; making them difficult and expensive to produce...
April 7, 2018: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/29563896/screening-of-a-scfv-antibody-with-high-affinity-for-application-in-human-ifn-%C3%AE-immunoassay
#8
Hang Yang, Yanfang Zhong, Juncheng Wang, Qinghong Zhang, Xiulan Li, Sumei Ling, Shihua Wang, Rongzhi Wang
Interferon gamma (IFN-γ), a signal proinflammatory cytokine secreted by immune cell, and plays a critical role in the pathogenesis and progression of many diseases. It has been regarded as an important marker for determination of disease-specific immune responses. Therefore, it is urgent to develop a feasible and accurate method to detect IFN-γ in clinic real blood samples. Until now, the immunoassay based on singe chain variable fragment (scFv) antibody for human IFN-γ is still not reported. In the present study, an scFv antibody named scFv-A8 with high specificity was obtained by phage display and biopanning, with the affinity 2...
2018: Frontiers in Microbiology
https://www.readbyqxmd.com/read/29545792/coupling-of-single-molecule-long-read-sequencing-with-imgt-highv-quest-analysis-expedites-identification-of-siv-gp140-specific-antibodies-from-scfv-phage-display-libraries
#9
Seung Yub Han, Alesia Antoine, David Howard, Bryant Chang, Woo Sung Chang, Matthew Slein, Gintaras Deikus, Sofia Kossida, Patrice Duroux, Marie-Paule Lefranc, Robert P Sebra, Melissa L Smith, Ismael Ben F Fofana
The simian immunodeficiency virus (SIV)/macaque model of human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome pathogenesis is critical for furthering our understanding of the role of antibody responses in the prevention of HIV infection, and will only increase in importance as macaque immunoglobulin (IG) gene databases are expanded. We have previously reported the construction of a phage display library from a SIV-infected rhesus macaque ( Macaca mulatta ) using oligonucleotide primers based on human IG gene sequences...
2018: Frontiers in Immunology
https://www.readbyqxmd.com/read/29484477/screening-for-single-chain-variable-fragment-antibodies-against-multiple-cry1-toxins-from-an-immunized-mouse-phage-display-antibody-library
#10
Sa Dong, Zongyi Bo, Cunzheng Zhang, Jianguo Feng, Xianjin Liu
Single-chain variable fragment (scFv) is a kind of antibody that possess only one chain of the complete antibody while maintaining the antigen-specific binding abilities and can be expressed in prokaryotic system. In this study, scFvs against Cry1 toxins were screened out from an immunized mouse phage displayed antibody library, which was successfully constructed with capacity of 6.25 × 107  CFU/mL. Using the mixed and alternative antigen coating strategy and after four rounds of affinity screening, seven positive phage-scFvs against Cry1 toxins were selected and characterized...
April 2018: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/29427674/phage-display-antibodies-against-ectromelia-virus-that-neutralize-variola-virus-selection-and-implementation-for-p35-neutralizing-epitope-mapping
#11
Yana Khlusevich, Andrey Matveev, Ivan Baykov, Leonid Bulychev, Nikolai Bormotov, Ivan Ilyichev, Georgiy Shevelev, Vera Morozova, Dmitrii Pyshnyi, Nina Tikunova
In this study, five phage display antibodies (pdAbs) against ectromelia virus (ECTV) were selected from vaccinia virus (VACV)-immune phage-display library of human single chain variable fragments (scFv). ELISA demonstrated that selected pdAbs could recognize ECTV, VACV, and cowpox virus (CPXV). Atomic force microscopy visualized binding of the pdAbs to VACV. Three of the selected pdAbs neutralized variola virus (VARV) in the plaque reduction neutralization test. Western blot analysis of ECTV, VARV, VACV, and CPXV proteins indicated that neutralizing pdAbs bound orthopoxvirus 35 kDa proteins, which are encoded by the open reading frames orthologous to the ORF H3L in VACV...
April 2018: Antiviral Research
https://www.readbyqxmd.com/read/29420662/generation-of-high-affinity-internalizing-anti-fgfr2-single-chain-variable-antibody-fragment-fused-with-fc-for-targeting-gastrointestinal-cancers
#12
Aleksandra Borek, Aleksandra Sokolowska-Wedzina, Grzegorz Chodaczek, Jacek Otlewski
Fibroblast growth factor receptors (FGFRs) are promising targets for antibody-based cancer therapies, as their substantial overexpression has been found in various tumor cells. Aberrant activation of FGF receptor 2 (FGFR2) signaling through overexpression of FGFR2 and/or its ligands, mutations, or receptor amplification has been reported in multiple cancer types, including gastric, colorectal, endometrial, ovarian, breast and lung cancer. In this paper, we describe application of the phage display technology to produce a panel of high affinity single chain variable antibody fragments (scFvs) against the extracellular ligand-binding domain of FGFR2 (ECD_FGFR2)...
2018: PloS One
https://www.readbyqxmd.com/read/29420566/a-novel-monoclonal-antibody-targeting-carboxymethyllysine-an-advanced-glycation-end-product-in-atherosclerosis-and-pancreatic-cancer
#13
Ulrika Wendel, Nina Persson, Christian Risinger, Eva Bengtsson, Björn Nodin, Lena Danielsson, Charlotte Welinder, Gunilla Nordin Fredrikson, Bo Jansson, Ola Blixt
Advanced glycation end products are formed by non-enzymatic reactions between proteins and carbohydrates, causing irreversible lysine and arginine alterations that severely affect protein structure and function. The resulting modifications induce inflammation by binding to scavenger receptors. An increase in advanced glycation end products is observed in a number of diseases e.g. atherosclerosis and cancer. Since advanced glycation end products also are present in healthy individuals, their detection and quantification are of great importance for usage as potential biomarkers...
2018: PloS One
https://www.readbyqxmd.com/read/29356992/identification-and-production-of-mouse-scfv-to-specific-epitope-of-enterovirus-71-virion-protein-2-vp2
#14
Jeeraphong Thanongsaksrikul, Potjanee Srimanote, Pongsri Tongtawe, Kittirat Glab-Ampai, Aijaz Ahmad Malik, Oratai Supasorn, Phatcharaporn Chiawwit, Yong Poovorawan, Wanpen Chaicumpa
Enterovirus-71 (EV71) and coxsackievirus-A16 (CA16) frequently cause hand-foot-mouth disease (HFMD) epidemics among infants and young children. CA16 infections are usually mild, while EV71 disease may be fatal due to neurologic complications. As such, the ability to rapidly and specifically recognize EV71 is needed to facilitate proper case management and epidemic control. Accordingly, the aim of this study was to generate antibodies to EV71-virion protein-2 (VP2) by phage display technology for further use in specific detection of EV71...
May 2018: Archives of Virology
https://www.readbyqxmd.com/read/29351623/development-of-sugar-chain-binding-single-chain-variable-fragment-antibody-to-adult-t-cell-leukemia-cells-using-glyco-nanotechnology-and-phage-display-method
#15
Kaname Muchima, Taro Todaka, Hiroyuki Shinchi, Ayaka Sato, Arisa Tazoe, Rikiya Aramaki, Yuhei Kakitsubata, Risa Yokoyama, Naomichi Arima, Masanori Baba, Masahiro Wakao, Yuji Ito, Yasuo Suda
Adult T-cell leukemia (ATL) is an intractable blood cancer caused by the infection of human T-cell leukemia virus type-1, and effective medical treatment is required. It is known that the structure and expression levels of cell surface sugar chains vary depending on cell states such as inflammation and cancer. Thus, it is expected that the antibody specific for ATL cell surface sugar chain would be an effective diagnostic tool and a strong candidate for the development of an anti-ATL drug. Here, we developed a stable sugar chain-binding single-chain variable fragment antibody (scFv) that can bind to ATL cells using a fibre-type Sugar Chip and phage display method...
April 1, 2018: Journal of Biochemistry
https://www.readbyqxmd.com/read/29330770/decoding-selection-bias-imparted-by-unpaired-cysteines-a-tug-of-war-between-expression-and-affinity
#16
B Vijayalakshmi Ayyar, Stephen Hearty, Richard O'Kennedy
In a recombinant antibody scFv format, the presence of an unpaired cysteine (Cys) is implicated in reduced soluble expression and inefficient presentation in phage display. Compared to other species, antibodies derived from rabbits are more likely to contain this unpaired Cys residue at position 80 (Cys80), when generated in a scFv format. In a screening campaign to isolate rabbit scFv against cardiac troponin I (cTnI), it was found that, a large proportion of isolated cTnI-specific clones contained unpaired Cys80...
January 12, 2018: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/29315611/isolation-of-a-high-affinity-bet-v-1-specific-igg-derived-scfv-from-a-subject-vaccinated-with-hypoallergenic-bet-v-1-fragments
#17
E Gadermaier, K Marth, C Lupinek, R Campana, G Hofer, K Blatt, D Smiljkovic, U Roder, M Focke-Tejkl, S Vrtala, W Keller, P Valent, R Valenta, S Flicker
BACKGROUND: Recombinant hypoallergenic allergen derivatives have been used in clinical immunotherapy studies, and clinical efficacy seems to be related to the induction of blocking IgG antibodies recognizing the wild-type allergens. However, so far no treatment-induced IgG antibodies have been characterized. OBJECTIVE: To clone, express, and characterize IgG antibodies induced by vaccination with two hypoallergenic recombinant fragments of the major birch pollen allergen, Bet v 1 in a nonallergic subject...
January 9, 2018: Allergy
https://www.readbyqxmd.com/read/29306173/blocking-osteopontin-fibronectin-interactions-reduce-extracellular-fibronectin-deployment-and-arthritic-immunopathology
#18
Brij Bhushan Mehta, Saniya Sharma, Rakesh K Vasishta, Ramesh K Sen, Aman Sharma, Manni Luthra-Guptasarma
Elevated levels of a thrombin-cleaved fragment of osteopontin (OPNT) are seen in synovial fluid (SF) and tissues of rheumatoid arthritis (RA) patients. OPNT binds to integrins on cell surfaces, inducing adhesion, migration and survival of inflammatory cells in the synovial joints, where OPNT binds to fibronectin to link fibroblast-like synoviocytes (FLS) with B cells, stimulating the latter to produce inflammatory cytokines. Our aim was to block OPNT-fibronectin interactions and examine whether this reduces inflammation...
February 2018: International Immunopharmacology
https://www.readbyqxmd.com/read/29305748/production-of-human-single-chain-fragment-antibody-scfv-against-human-epidermal-growth-factor-receptor-2-her-2-by-phage-display-technology
#19
Saeideh Foroumadi, Masoumeh Rajabibazl, Azam Rahimpour, Solmaz Shahidi, Walead Ebrahimizadeh, Maral Yarahmadi, Shirin Rajabi, Azam Daraei
Breast cancer with more than 1.7 million diagnoses per year has been known as one of the most prevalent cancers among women worldwide. Despite the availability of advanced treatment options, cancer progression and metastasis is observed in 20% of patients. Human epidermal growth factor receptor-2 (HER-2) is considered as an important prognostic and diagnostic tumor marker for breast cancer. While HER-2 is expressed on the surface of normal cells, its overexpression occurs in 20-25% on breast cancer tumor cells...
February 2018: In Vitro Cellular & Developmental Biology. Animal
https://www.readbyqxmd.com/read/29199242/a-single-step-breeding-generated-a-diagnostic-anti-cortisol-antibody-fragment-with-over-30-fold-enhanced-affinity
#20
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
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