Recombinant human antibody fragment against tetanus toxoid produced by phage display

Neelakantam, B.; Sridevi, N.; Shukra, A. M.; Sugumar, Parthsarathy; Samuel, Sherin; Lingala, Rajendra

doi:10.1556/eujmi.4.2014.1.4

Cited by 10 publications

(4 citation statements)

References 25 publications

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“…Numerous monoclonal antibodies have been developed against TeNT and tested for their neutralizing activity. Some monoclonal antibodies were generated against TeNT using mouse hybridoma technology [ 29 , 30 , 31 ], and some were selected from human phage libraries [ 32 , 33 ]. However, there are few existing reports describing selection from human immune libraries.…”

Section: Discussionmentioning

confidence: 99%

Tetanus Neurotoxin Neutralizing Antibodies Screened from a Human Immune scFv Antibody Phage Display Library

Wang

Fang

et al. 2016

Toxins

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Tetanus neurotoxin (TeNT) produced by Clostridium tetani is one of the most poisonous protein substances. Neutralizing antibodies against TeNT can effectively prevent and cure toxicosis. Using purified Hc fragments of TeNT (TeNT-Hc) as an antigen, three specific neutralizing antibody clones recognizing different epitopes were selected from a human immune scFv antibody phage display library. The three antibodies (2-7G, 2-2D, and S-4-7H) can effectively inhibit the binding between TeNT-Hc and differentiated PC-12 cells in vitro. Moreover, 2-7G inhibited TeNT-Hc binding to the receptor via carbohydrate-binding sites of the W pocket while 2-2D and S-4-7H inhibited binding of the R pocket. Although no single mAb completely protected mice from the toxin, they could both prolong survival when challenged with 20 LD50s (50% of the lethal dose) of TeNT. When used together, the mAbs completely neutralized 1000 LD50s/mg Ab, indicating their high neutralizing potency in vivo. Antibodies recognizing different carbohydrate-binding pockets could have higher synergistic toxin neutralization activities than those that recognize the same pockets. These results could lead to further production of neutralizing antibody drugs against TeNT and indicate that using TeNT-Hc as an antigen for screening human antibodies for TeNT intoxication therapy from human immune antibody library was convenient and effective.

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Section: Discussionmentioning

confidence: 99%

Tetanus Neurotoxin Neutralizing Antibodies Screened from a Human Immune scFv Antibody Phage Display Library

Wang

Fang

et al. 2016

Toxins

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“…Phages are passed down a column containing the target of interest, which binds phages containing an active peptide motif. This technique has been employed for the identification of affinity 31,32 or avidity binding proteins for antibody-antigen interactions 31,33 and sequences that inhibit PPIs. 34 Phages displaying inhibitory proteins are then amplified and reintroduced to the column under more stringent conditions (e.g., fewer binding sites, competitive inhibitor; known as “biopanning”).…”

Section: Phage/phagemid Displaymentioning

confidence: 99%

Methods for the Creation of Cyclic Peptide Libraries for Use in Lead Discovery

et al. 2015

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A surprisingly high number of drug discovery projects begin with a peptide as the initial hit.1-4 These starting molecules typically need significant chemical development as linear peptides have poor pharmacokinetic properties (e.g., oral bioavailability and stability to peptidases). 5,6 Cyclic peptides, however, are far less susceptible to proteolysis 7 and often have increased biological activity because of their conformational rigidity, which decreases entropic loss upon binding. 8,9 In contrast to (and possibly as a result of) their relative underutilization in industry, cyclic peptide libraries have been extensively employed in academia, with multiple approaches developed for their generation. Such work has demonstrated the suitability of the cyclic peptide scaffold for the identification of inhibitors against some of the most challenging targets, including protein-protein interactions (PPIs).10 Macrocyclic peptide scaffolds appear to be optimal for this purpose, and with genetically encoded cyclic peptide libraries in particular, 11,12 there is potential for the straightforward creation of large sequence diversity (e.g., 6.4 × 10 7 members for six randomized amino acids). For synthetic cyclic peptide libraries, there are several approaches for identification of hits, including mass spectrometry, or by sequencing an associated genetic tag (incorporated during library synthesis). 13For the purpose of this review, we have divided cyclic peptide libraries into three main categories: biologic, semisynthetic, and fully synthetic. Some of these libraries are constrained to the canonical peptidogenic amino acids, whereas others are able to include nonpeptidogenic amino acids such as β-or γ-amino acids, D-amino acids, or nonnatural amino acids. 14-16The majority of biologically produced cyclic peptide libraries are formed using phage/phagemid display 8,17 or by splitintein cyclisation of peptides and proteins (SICLOPPS). 11,18,19 The latter uses a selectively randomized library of splitinteins for the production of genetically encoded, backbone cyclized peptide libraries. Semisynthetic libraries are able to incorporate nonpeptidogenic amino acids while retaining ribosomal synthesis and an associated genetic tag. The most frequently used method for semisynthetic library production is mRNA display 7,20 ; novel variants thereof include random nonstandard peptide integrated discovery (RaPID) 14 or protein synthesis using recombinant elements (PURE).15 Both methods use promiscuous enzymes to expand the amino acid library encoded into peptides by reprogramming codons. 16Another semisynthetic technique has been used for the creation of macrocyclic organic-peptide hybrids (MOrPHs) 21,22 and bicyclic organo-peptide hybrids (BOrPHs), 23 which produce geometrically constrained peptide libraries. Fully AbstractThe identification of initial hits is a crucial stage in the drug discovery process. Although many projects adopt high-throughput screening of small-molecule libraries at this stage, there is significant potential for s...

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“…Phage library panning experiments against C. difficile toxin B generated a mAb, with improved affinity over the parent antibody, intended for use as a novel immunodiagnostic agent (97). Clostridium tetani, another member of this Clostridium genus, (98). While the majority of successful phage library panning campaigns involve exotoxins, less successful experiments involving bacterial surface proteins are described in the literature.…”

Section: Other Antibacterial Mabs Discovered By Phage Displaymentioning

confidence: 99%

Phage and Yeast Display

Sheehan

Marasco

2015

Microbiol Spectr

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Despite the availability of antimicrobial drugs, the continued development of microbial resistance-established through escape mutations and the emergence of resistant strains-limits their clinical utility. The discovery of novel, therapeutic, monoclonal antibodies (mAbs) offers viable clinical alternatives in the treatment and prophylaxis of infectious diseases. Human mAb-based therapies are typically nontoxic in patients and demonstrate high specificity for the intended microbial target. This specificity prevents negative impacts on the patient microbiome and avoids driving the resistance of nontarget species. The in vitro selection of human antibody fragment libraries displayed on phage or yeast surfaces represents a group of well-established technologies capable of generating human mAbs. The advantage of these forms of microbial display is the large repertoire of human antibody fragments present during a single selection campaign. Furthermore, the in vitro selection environments of microbial surface display allow for the rapid isolation of antibodies-and their encoding genes-against infectious pathogens and their toxins that are impractical within in vivo systems, such as murine hybridomas. This article focuses on the technologies of phage display and yeast display, as these strategies relate to the discovery of human mAbs for the treatment and vaccine development of infectious diseases.

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Recombinant human antibody fragment against tetanus toxoid produced by phage display

Cited by 10 publications

References 25 publications

Tetanus Neurotoxin Neutralizing Antibodies Screened from a Human Immune scFv Antibody Phage Display Library

Tetanus Neurotoxin Neutralizing Antibodies Screened from a Human Immune scFv Antibody Phage Display Library

Methods for the Creation of Cyclic Peptide Libraries for Use in Lead Discovery

Phage and Yeast Display

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