Multiplex epitope mapping using bacterial surface display reveals both linear and conformational epitopes

Hudson, Elton P.; Uhlén, Mathias; Rockberg, Johan

doi:10.1038/srep00706

Cited by 26 publications

(21 citation statements)

References 34 publications

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“…As an alternative, pAbs are produced as ascites in mice. Moreover, these antibodies have superior specificity compared with monoclonal antibodies because they are generated by a large number of B-cell clones each producing antibodies to a specific epitope (Hudson et al, 2012;Zhuang et al, 2014).…”

Section: Polyclonal Antibodymentioning

confidence: 99%

Antibody Engineering for Pursuing a Healthier Future

et al. 2017

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Since the development of antibody-production techniques, a number of immunoglobulins have been developed on a large scale using conventional methods. Hybridoma technology opened a new horizon in the production of antibodies against target antigens of infectious pathogens, malignant diseases including autoimmune disorders, and numerous potent toxins. However, these clinical humanized or chimeric murine antibodies have several limitations and complexities. Therefore, to overcome these difficulties, recent advances in genetic engineering techniques and phage display technique have allowed the production of highly specific recombinant antibodies. These engineered antibodies have been constructed in the hunt for novel therapeutic drugs equipped with enhanced immunoprotective abilities, such as engaging immune effector functions, effective development of fusion proteins, efficient tumor and tissue penetration, and high-affinity antibodies directed against conserved targets. Advanced antibody engineering techniques have extensive applications in the fields of immunology, biotechnology, diagnostics, and therapeutic medicines. However, there is limited knowledge regarding dynamic antibody development approaches. Therefore, this review extends beyond our understanding of conventional polyclonal and monoclonal antibodies. Furthermore, recent advances in antibody engineering techniques together with antibody fragments, display technologies, immunomodulation, and broad applications of antibodies are discussed to enhance innovative antibody production in pursuit of a healthier future for humans.

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Section: Polyclonal Antibodymentioning

confidence: 99%

Antibody Engineering for Pursuing a Healthier Future

et al. 2017

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“…Several approaches combining the use of computational analysis with laboratory techniques have been widely described in the scientific literature [88][89][90][91][92][93]. Here we take influenza virus as an example of hypervariable pathogen that requires the development of novel vaccinal strategies to elicit a broad immune response.…”

Section: Discussionmentioning

confidence: 99%

Peptide-Based Vaccinology: Experimental and Computational Approaches to Target Hypervariable Viruses through the Fine Characterization of Protective Epitopes Recognized by Monoclonal Antibodies and the Identification of T-Cell-Activating Peptides

Castelli

Cappelletti

Diotti

et al. 2013

Clinical and Developmental Immunology

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Defining immunogenic domains of viral proteins capable of eliciting a protective immune response is crucial in the development of novel epitope-based prophylactic strategies. This is particularly important for the selective targeting of conserved regions shared among hypervariable viruses. Studying postinfection and postimmunization sera, as well as cloning and characterization of monoclonal antibodies (mAbs), still represents the best approach to identify protective epitopes. In particular, a protective mAb directed against conserved regions can play a key role in immunogen design and in human therapy as well. Experimental approaches aiming to characterize protective mAb epitopes or to identify T-cell-activating peptides are often burdened by technical limitations and can require long time to be correctly addressed. Thus, in the last decade many epitope predictive algorithms have been developed. These algorithms are continually evolving, and their use to address the empirical research is widely increasing. Here, we review several strategies based on experimental techniques alone or addressed by in silico analysis that are frequently used to predict immunogens to be included in novel epitope-based vaccine approaches. We will list the main strategies aiming to design a new vaccine preparation conferring the protection of a neutralizing mAb combined with an effective cell-mediated response.

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“…The display of complementary affibodies was also assessed for enabling binding between Synechocystis and the heterotrophic bacterium E. coli or S. carnosus. Both of these bacteria have established surface display systems: E. coli display is based on the adhesin involved in diffuse adherence (AIDA-I) autotransporter (45), while S. carnosus display is based on the cell wall-anchoring region of Staphylococcus aureus protein A (43,51). The E. coli and S. carnosus display constructs contain the streptococcal protein G albumin binding domain (ABD) (5.1 kDa) and albumin binding protein (ABP) (21.8 kDa) as spacers, respectively (52).…”

Section: Display Of Polymerizing Affibodies For Affinity-drivenmentioning

confidence: 99%

Surface Display of Small Affinity Proteins on Synechocystis sp. Strain PCC 6803 Mediated by Fusion to the Major Type IV Pilin PilA1

2018

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Functional surface display of small affinity proteins, namely Affibodies (6.5 kDa), was evaluated for the model cyanobacterium sp. PCC 6803, through anchoring to native surface structures. These structures included confirmed or putative subunits of the type IV pili, the S-layer protein, and the heterologous autotransporter antigen 43 system. The most stable display system was determined to be through C-terminal fusion to PilA1, the major type IV pili subunit in , in a strain unable to retract these pili (Δ). Type IV pili synthesis was upheld, albeit reduced, when fusion proteins were incorporated. However, pili-mediated functions such as motility and transformational competency were negatively affected. Display of Affibodies on and the complementary anti-idiotypic Affibodies on or was able to mediate interspecies cell-cell binding by Affibody complex formation. The same strategy was however not able to drive cell-cell binding and aggregation of-only mixtures. Successful Affibody-tagging of the putative minor pilin PilA4 showed that it locates to the type IV pili in , and that its extracellular availability depends on PilA1. In addition, Affibody-tagging of the S-layer protein indicated that the domains responsible for anchoring and secretion of this protein are located at the N- and C-terminus, respectively. This study can serve as a basis for future surface display of proteins on for biotechnological applications. Cyanobacteria are gaining interest for their potential as autotrophic cell factories. Development of efficient surface display strategies could improve their suitability for large-scale applications by providing options for designed microbial consortia, cell immobilization, and biomass harvesting. Here, surface display of small affinity proteins was realized by fusing them to the major subunit of the native type IV pili in sp. PCC 6803. Display of complementary affinity proteins allowed specific cell-cell binding between and or Additionally, successful tagging of the putative pilin PilA4, helped determine its localization to the type IV pili. Analogous tagging of the S-layer protein shed light on the regions involved in its secretion and surface anchoring.

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Multiplex epitope mapping using bacterial surface display reveals both linear and conformational epitopes

Cited by 26 publications

References 34 publications

Antibody Engineering for Pursuing a Healthier Future

Antibody Engineering for Pursuing a Healthier Future

Peptide-Based Vaccinology: Experimental and Computational Approaches to Target Hypervariable Viruses through the Fine Characterization of Protective Epitopes Recognized by Monoclonal Antibodies and the Identification of T-Cell-Activating Peptides

Surface Display of Small Affinity Proteins on Synechocystis sp. Strain PCC 6803 Mediated by Fusion to the Major Type IV Pilin PilA1

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