Related Experiment Video
Updated: May 12, 2026
09:06
Efficient and Site-specific Antibody Labeling by Strain-promoted Azide-alkyne Cycloaddition
Published on: December 23, 2016
20.7K
Binding mode-guided development of high-performance antibodies targeting site-specific posttranslational
Mariapia Riso1, Rohan N Shah2,3, Akiko Koide1,4
1Laura and Isaac Perlmutter Cancer Center, New York University Langone Health, New York, NY 10016.
Summary
Researchers developed a new platform for creating highly specific recombinant antibodies targeting protein posttranslational modifications (PTMs). This antigen clasping method ensures molecularly defined and renewable antibodies, improving reproducibility in biomedical research.
Area of Science:
- Biochemistry
- Molecular Biology
- Epigenetics
Background:
- Posttranslational modifications (PTMs) regulate crucial cellular events.
- Generating highly specific antibodies for PTMs is challenging due to difficulties in molecular recognition.
- Current antibodies can be inconsistent and nonrenewable, impacting research reproducibility.
Purpose of the Study:
- To develop a platform for generating potent, selective, and renewable recombinant antibodies to PTMs.
- To leverage the 'antigen clasping' binding mode for enhanced antibody performance.
- To address the reproducibility challenges in biomedical research linked to antibody variability.
Main Methods:
- Utilized a binding mode-guided approach based on the 'antigen clasping' mechanism.
- Designed a platform to generate clasping antibodies with two distinct binding units.
- Applied the platform to generate antibodies against trimethylated histone H3 and phosphotyrosine antigens.
Main Results:
- Successfully generated potent and selective recombinant antibodies to PTMs.
- Clasping antibodies demonstrated high specificity and affinity for trimethylated histone H3.
- A clasping antibody to H3K27me3 showed superior specificity in chromatin immunoprecipitation compared to conventional antibodies.
- The platform efficiently generated clasping antibodies to phosphotyrosine antigens.
Conclusions:
- The developed platform efficiently generates molecularly defined and renewable clasping antibodies for PTMs.
- Antigen clasping antibodies offer superior specificity and affinity, enhancing PTM detection and enrichment.
- This platform has broad applicability for generating high-performance antibodies against diverse PTMs, improving research reliability.
More Related Videos
11:02Genetic Encoding of a Non-Canonical Amino Acid for the Generation of Antibody-Drug Conjugates Through a Fast Bioorthogonal Reaction
Published on: September 14, 2018
7.7K
08:47Synthesis and Bioconjugation of Thiol-Reactive Reagents for the Creation of Site-Selectively Modified Immunoconjugates
Published on: March 6, 2019
9.4K
Related Concept Videos
01:49Conserved Binding Sites
Many proteins’ biological role depends on their interactions with their ligands, small molecules that bind to specific locations on the protein known as ligand-binding sites. Ligand-binding sites are often conserved among homologous proteins as these sites are critical for protein function.
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally analyses the...
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally analyses the...
02:07Improving Translational Accuracy
Base complementarity between the three base pairs of mRNA codon and the tRNA anticodon is not a failsafe mechanism. Inaccuracies can range from a single mismatch to no correct base pairing at all. The free energy difference between the correct and nearly correct base pairs can be as small as 3 kcal/ mol. With complementarity being the only proofreading step, the estimated error frequency would be one wrong amino acid in every 100 amino acids incorporated. However, error frequencies observed in...
02:28Leaky Scanning
During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA. Marilyn Kozak discovered that the sequence RCCAUGG (where R stands for...
02:07Improving Translational Accuracy
Base complementarity between the three base pairs of mRNA codon and the tRNA anticodon is not a failsafe mechanism. Inaccuracies can range from a single mismatch to no correct base pairing at all. The free energy difference between the correct and nearly correct base pairs can be as small as 3 kcal/ mol. With complementarity being the only proofreading step, the estimated error frequency would be one wrong amino acid in every 100 amino acids incorporated. However, error frequencies observed in...
01:20Immunoprecipitation
Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...
01:20Immunogold Electron Microscopy
Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.