Production and Screening of Monoclonal Peptide Antibodies

Trier, Nicole Hartwig; Mortensen, Anne S.; Schiolborg, Annette; Friis, T

doi:10.1007/978-1-4939-2999-3_12

Cited by 8 publications

(24 citation statements)

References 38 publications

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“…Similar results have been obtained with many other MAbs, which verifies the stringent requirements for an optimal antibody-antigen interaction [33,46,96,97]. MAbs may be used as crude culture supernatants (which usually contain bovine Ig) or after purification, e.g., on protein A/G [94,98] or synthetic resins, as illustrated by Islam et al in this issue [90]. However, it is important to remember, that such purification methods may not remove the bovine IgG, which originates from the cell culture medium [99,100].…”

Section: Monoclonal Antibodiessupporting

confidence: 80%

“…Köhler and Milstein pioneered the technique for producing monoclonal Abs (MAbs) of desired specificities [92,93]. The initial steps for generation of PAbs and MAbs are similar [86,87,94]. The immunogen is injected into the host along with an adjuvant, which is followed by repetitive immunizations to boost the immune response of the immunized animal.…”

Section: Monoclonal Antibodiesmentioning

confidence: 99%

“…As described above, the Abs generated are polyclonal, and, if aiming for MAbs, isolated B cells from the immunized animal are fused with myeloma cells. Single clones producing a sufficient amount of Abs are identified by screening and expansion [94]. The main advantage of isolating clones is that MAbs may be produced in unlimited quantities.…”

Section: Monoclonal Antibodiesmentioning

confidence: 99%

“…The main advantage of isolating clones is that MAbs may be produced in unlimited quantities. This technique is still the major method for MAb production, as described by Köhler and Milstein, even though minor modifications may be introduced, mainly in the screening phase, where an assay system identical to the end stage use should be used to avoid selection of clones with aberrant properties [94]. Upon successful screening and cloning, highly specific MAbs are obtained and the epitopes of these can be characterised with high precision, as illustrated by recent studies of MAbs to the glutamate decarboxylase isoforms 65 and 67, which were shown to recognise linear epitopes located in flexible structures [47,95].…”

Section: Monoclonal Antibodiesmentioning

confidence: 99%

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Peptides, Antibodies, Peptide Antibodies and More

Trier

Hansen

Houen

2019

IJMS

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The applications of peptides and antibodies to multiple targets have emerged as powerful tools in research, diagnostics, vaccine development, and therapeutics. Antibodies are unique since they, in theory, can be directed to any desired target, which illustrates their versatile nature and broad spectrum of use as illustrated by numerous applications of peptide antibodies. In recent years, due to the inherent limitations such as size and physical properties of antibodies, it has been attempted to generate new molecular compounds with equally high specificity and affinity, albeit with relatively low success. Based on this, peptides, antibodies, and peptide antibodies have established their importance and remain crucial reagents in molecular biology.

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Section: Monoclonal Antibodiessupporting

confidence: 80%

Section: Monoclonal Antibodiesmentioning

confidence: 99%

Section: Monoclonal Antibodiesmentioning

confidence: 99%

Section: Monoclonal Antibodiesmentioning

confidence: 99%

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Peptides, Antibodies, Peptide Antibodies and More

Trier

Hansen

Houen

2019

IJMS

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“…The concentrations of purified ascites were determined by Bradford method. The class and subclass of mAbs were determined by immunoType ™ mouse monoclonal antibody isotyping kit (Thermo Fisher, Waltham, MA, USA, Cat#26178) [18][19][20]. Following this process and primary screen, we obtained monoclonal antibodies against ZEBOV GP or VP40, which summarized in Table 1.…”

Section: Design and Production Of Mabsmentioning

confidence: 99%

Development and Characterization of Neutralizing Antibodies Against Zaire Ebolavirus Glycoprotein and Protein 40

Weng

Shen

et al. 2018

Cell Physiol Biochem

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Background/Aims: Monoclonal antibodies (mAbs) are presently the most promising treatment against Ebola virus disease (EVD), and cocktail of two or more antibodies likely confers protection through complementary mechanisms. Zaire Ebolavirus (EBOV) glycoprotein (GP) and viral protein 40 (VP40) are targets for designing neutralizing antibodies. Currently, the antiviral therapeutics of mAb-cocktails are still limited solely to anti-GP antibodies，there is no Abs cocktail against Zaire EBOV GP and VP40, which both have important interactions with host cellular membrane. Methods: We used hybridoma technology to produce anti-Zaire EBOV GP mAb against GP receptor binding domain, and anti-Zaire EBOV VP40 mAbs against the N-terminal domain, the C-terminal domain, respectively; synthesized Zaire EBOV transcription and replication competent virus like particles (trVLPs), which model even all aspects of the EBOV life cycles in order to evaluate the anti-viral effect of mAbs. Then, we characterized the anti- Zaire EBOV trVLPs effect of anti-GP and VP40 mAbs in vitro by real time-PCR, immunofluorescence assay and western blot analysis. Results: Our results demonstrate that anti-GP or anti-VP40 mAbs effectively inhibit trVLPs replication. The cocktails of anti-GP and anti-VP40 mAbs, or between anti-VP40 mAbs, had synergistic anti-trVLPs effect. Meanwhile, the detailed DNA and amino acid sequences of the mAbs were checked. Conclusion: The study verifies neutralizing efficacy of anti-GP or anti-VP40 mAb, report promising cocktail of anti-GP and anti-VP40 mAb, or cocktail of two anti-VP40 mAbs. To our knowledge, this is the first account to report the important anti-viral effect of cocktails of anti-GP and anti-VP40 mAbs in vitro.

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Production of Antibodies to Peptide Targets Using Hybridoma Technology

Trier,

Friis

2024

Methods in Molecular Biology

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Production and Screening of Monoclonal Peptide Antibodies

Cited by 8 publications

References 38 publications

Peptides, Antibodies, Peptide Antibodies and More

Peptides, Antibodies, Peptide Antibodies and More

Development and Characterization of Neutralizing Antibodies Against Zaire Ebolavirus Glycoprotein and Protein 40

Production of Antibodies to Peptide Targets Using Hybridoma Technology

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