Related Experiment Video
Updated: Jan 31, 2026

Activated Cross-linked Agarose for the Rapid Development of Affinity Chromatography Resins - Antibody Capture as a Case Study
Published on: August 16, 2019
Development, Optimization, and Structural Characterization of an Efficient Peptide-Based Photoaffinity Cross-Linking
Nicholas Vance1, Neelie Zacharias1, Mark Ultsch1
1Research & Early Development , Genentech, Inc. , 1 DNA Way , South San Francisco , California 94080 , United States.
A new photoaffinity cross-linking method enables site-specific conjugation of molecules to antibodies without genetic modification. This approach simplifies antibody conjugate preparation and maintains biological activity for targeted therapies.
Area of Science:
- Bioconjugation Chemistry
- Antibody Engineering
- Drug Development
Background:
- Site-specific antibody conjugation is crucial for developing homogeneous targeted therapies and diagnostics.
- Current methods often require antibody glycan or disulfide bond modification, complicating preparation and potentially affecting biological activity.
Purpose of the Study:
- To develop a robust photoaffinity cross-linking method for site-specific conjugation to wild-type antibodies.
- To characterize the method's efficiency, scope, and the precise site of conjugation.
- To demonstrate the utility of the method for creating antibody-drug conjugates.
Main Methods:
- Utilized a benzoylphenylalanine (Bpa) mutant peptide that binds the antibody Fc domain.
- Employed UV irradiation to activate Bpa, forming a diradical for cross-linking to the antibody.
- Characterized conjugation site using mass spectrometry and X-ray crystallography.
- Adapted the method to conjugate a cytotoxic payload to an antibody.
Main Results:
- Developed an efficient photoaffinity cross-linking method for site-specific conjugation to glycosylated wild-type antibodies.
- Confirmed efficient conjugation without UV-induced antibody photodamage.
- Identified the specific conjugation site on a human antibody.
- Created a stable and potent antibody-drug conjugate with potential for therapeutic applications.
Conclusions:
- The developed photoaffinity cross-linking method offers a simplified and effective approach for site-specific antibody conjugation.
- This technique preserves antibody integrity and biological activity, paving the way for advanced antibody-based therapeutics and diagnostics.
More Related Videos
08:40Photo-Induced Cross-Linking of Unmodified Proteins PICUP Applied to Amyloidogenic Peptides
Published on: January 12, 2009
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
Related Concept Videos
Antibody Structure
Antibodies, also known as immunoglobulins (Ig), are essential players of the adaptive immune system. These antigen-binding proteins are produced by B cells and make up 20 percent of the total blood plasma by weight. In mammals, antibodies fall into five different classes, which each elicits a different biological response upon antigen binding.
The Y-Shaped Structure of Antibodies Consists of Four Polypeptide Chains
Antibodies consist of four polypeptide chains: two identical heavy...
Homogeneous Equilibria for Gaseous Reactions
For gas-phase reactions, the equilibrium constant may be expressed in terms of either the molar concentrations (Kc) or partial pressures (Kp) of the reactants and products. A relation between these two K values may be simply derived from the ideal gas equation and the definition of molarity. According to the ideal gas equation:
Crossed Aldol Reaction Using Weak Bases
Crossed Aldol Reaction Using Strong Bases: Directed Aldol Reaction
Peptide Bonds
Structure of Conjugated Dienes
Conjugated dienes are compounds characterized by the presence of alternating double and single bonds. In a conjugated system like 1,3-butadiene, the unhybridized 2p orbital on each carbon overlaps continuously, allowing the π electrons to be delocalized across the entire molecule. In contrast, this type of overlap does not occur in cumulated and isolated dienes, such as 2,3-pentadiene and 1,4-pentadiene, respectively. Instead, the π electrons remain localized between the double...