Epitope Mapping of Mono- and Polyclonal Antibodies by Screening Phage-displayed Random Peptide Libraries

Molek, Peter; Bratkovič, Tomaž

doi:10.17344/acsi.2016.2458

Cited by 7 publications

(4 citation statements)

References 25 publications

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“…Extract from noninfected bacteria and an unrelated pIII-fused peptide (linear GTFDHPQ targeting streptavidin) were used as controls. 27 Sera pool showed IgE binding to the six pIII-fused peptides (Fig. 5).…”

Section: Resultsmentioning

confidence: 98%

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Epitope Mapping of Major Ragweed Allergen Amb a 1

Zahirović

Štrukelj

Korošec

et al. 2019

ACSi

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Ragweed is a prominent cause of seasonal allergies. Thus far, information on IgE-binding sites of major allergen in ragweed pollen, Amb a 1, is very limited. A powerful experimental method to gain insights on the allergen epitopes is the selection of peptides from biological libraries that bind to anti-allergen antibodies. In this work, we aimed to map IgE epitopes of Amb a 1 using epitope-mimicking short peptides-mimotopes that were affinity-selected from phage-displayed random peptide libraries. The peptides weakly aligned with the Amb a 1 primary sequence, thus suggesting that the epitopes are conformational. When the peptides were mapped onto the surface of Amb a 1 homology model, the EpiSearch analysis predicted the location of four potential epitopic sites on surface patches centred at residues K 104 , S 110 , H 214 , and W 312. The peptides matching to the predicted epitopes bound selectively to the IgE from pool of ragweed-allergic patients' sera and therefore represent mimetics of Amb a 1 IgE epitopes. The knowledge of IgE epitopes is a prerequisite for the rational design of molecular-based approaches to diagnosis and immunotherapy of allergic diseases.

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Section: Resultsmentioning

confidence: 98%

“…Extracts of non-infected bacteria and bacteria infected with phage clone carrying unrelated pIII-fused control peptide (linear GTFDHPQ targeting streptavidin) were prepared in the same way and used as negative controls. 27…”

Section: Isolation Of Piii-fused Peptides From E Coli Periplasmmentioning

confidence: 99%

Epitope Mapping of Major Ragweed Allergen Amb a 1

Zahirović

Štrukelj

Korošec

et al. 2019

ACSi

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“…Consequently, the mapping of conformational epitopes is complicated and usually requires other ways to complete, such as computational tools and phage display techniques. These methods are much more difficult than traditional linear epitope identification methods [51] , [52] . Functional methods are used to detect the binding efficiency of antibody with protein fragments, synthetic peptides or recombinant proteins, and more convenient to conduct [50] .…”

Section: Discussionmentioning

confidence: 99%

Precise location of two novel linear epitopes on the receptor-binding domain surface of MERS-CoV spike protein recognized by two different monoclonal antibodies

Wang

Ding

Wei

et al. 2022

International Journal of Biological Macromolecules

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Middle East respiratory syndrome coronavirus (MERS-CoV) is a coronavirus which can cause severe human respiratory diseases with a fatality rate of almost 36%. In this study, we report the generation, characterization and epitope mapping of several monoclonal antibodies against the spike receptor-binding domain (RBD) of MERS-CoV. Two monoclonal antibodies (4C7 and 6E8) that can react with linearized RBD have been selected for subsequent identification of RBD mAb-binding epitopes. Two distinct novel linear epitopes, 423 FTCSQIS 429 and 546 SPLEGGGWL 554 ,were precisely located at the outermost surface of RBD by dot-blot hybridization and ELISAs. Multiple sequence alignment analysis showed that these two peptides were highly conserved. Alanine (A)-scanning mutagenesis demonstrated that residues 423F, 428I, and 429S are the crucial residues for the linear epitope 423 FTCSQIS 429 while residues 548L, 550G, 553W, 554L for epitope 546 SPLEGGGWL 554 . These findings may be helpful for further understanding of the function of RBD protein and the development of subsequent diagnosis and detection methods.

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“…HLA-A2, having the highest prevalence of MHC class I in the Thai population, was selected for this study. The MNP, Fe 3 O 4 , is considered due to its ability to direct a magnetic field, low toxicity, strong biocompatibility, and high relaxivity [16][17][18][19][20][21][22][23][24][25][26][27][28][29][30][31][32][33][34] in a biosensing system based on micro-NMR. Owing to the instability of iron in a strongly acidic environment, MNPs may shorten the lifespan of a material.…”

Section: Introductionmentioning

confidence: 99%

Immobilization of Thai Population-specific Human Leukocyte Antigens on Magnetic Nanoparticles Integrated with Nuclear Magnetic Resonance Technology

Subtaweewasin,

Pijitrojana

2023

Nano Biomedicine and Engineering

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Kidney disease, as a global health problem, can progress to kidney failure. Kidney transplantation is a treatment option for end-stage disease and an alternative to dialysis. Complement-dependent cytotoxicity tests and flow cytometry are methods used to test for compatibility between autoantibodies and donor recipient. Antibodies against donor human leukocyte antigens (HLAs), linked to hyperacute or severely acute rejection, frequently result in graft loss. Currently, fluorescent bead assay (Luminex ® ) technology is used in Thailand for HLA-antibody typing tests. However, each test requires specific location and time, expensive equipment, and high costs. The goal of such research is that, HLA-antibody typing tests can be performed wherever it is the most convenient for recipients and hospitals. A further advantage is that the surgeon can perform a final test for confirmation before the surgery. Numerous technologies are currently available for this purpose. The focus of this study was on the Fe 3 O 4 magnetic nanoparticles (MNPs) immobilized with HLAs frequently found in the Thai population. These were examined using nuclear magnetic resonance (NMR) to measure HLA antibody typing. MNPs have recently attracted considerable attention because of their strong magnetization and large surface areas. Immobilization reactions with antigens and the straightforward magnetic separation of MNPs are low-cost techniques. Hence, the commercialization of Fe 3 O 4 MNPs is favorable. In this study, MNPs were functionalized with 3aminopropyl triethoxysilane (APTES) in toluene and ethanol, and the nanoparticles were streptavidin activated after APTES treatment. A specific biotinylated HLA for the Thai population was used and immobilized on streptavidin. The MNPs were effectively immobilized. The magnetic properties can be modified by adding other ions to Fe 3 O 4 . We discovered that the shape and size variations of Fe 3 O 4 MNPs may have an impact on the amount of functionalized and immobilized surface area.

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Epitope Mapping of Mono- and Polyclonal Antibodies by Screening Phage-displayed Random Peptide Libraries

Cited by 7 publications

References 25 publications

Epitope Mapping of Major Ragweed Allergen Amb a 1

Epitope Mapping of Major Ragweed Allergen Amb a 1

Precise location of two novel linear epitopes on the receptor-binding domain surface of MERS-CoV spike protein recognized by two different monoclonal antibodies

Immobilization of Thai Population-specific Human Leukocyte Antigens on Magnetic Nanoparticles Integrated with Nuclear Magnetic Resonance Technology

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