Creative Proteomics offers conjugation position analysis service to identify specific sites or locations where conjugation reactions occur to form conjugated molecules with different properties and reactivity, helping clients worldwide to meet the requirements of ICH Q6B guidelines.
Background
The locus of conjugation, denoted as the conjugation position, pertains to the definitive nexus or site within a molecular structure where the process of conjugation transpires. The biological process of conjugation encompasses the amalgamation of a minimum of two molecules, laying down the groundwork for the formation of a novel compound or entity. In the realm of chemistry, the term conjugation signifies the process wherein addition of chemical functional groups or molecular substructures, known as moieties, to molecules transpire, resulting in the modification or alteration of its characteristics or reactivity. The conjugation position inherently exhibits variability contingent upon the particular molecules implicated and the kind of reaction that is underway.
Fig 1. Current LC-MS-based strategies for characterization and quantification of antibody-drug conjugates. (Zhu, X., et al.; 2020)
The significance of the conjugation position is underscored by its impact on the final structure and attributes of the new molecular compound that results from conjugation. To illustrate, within the scope of organic chemistry, conjugated systems are established when pi (π) bonds exist in neighboring positions along the backbone of a molecule, thereby enabling the delocalization of electrons. The geographical position of these conjugated pi bonds on the molecular landscape decides the electronic properties, which could include the molecule's ability to absorb light or engage in resonance stabilization.
What can we provide for conjugation position analysis?
Mass Spectrometry
In bioanalytical techniques, Mass Spectrometry (MS) is a robust tool utilized to ascertain protein conjugation sites. It operates through the ionization of protein molecules, followed by the measurement of their respective mass-to-charge ratios. Depending on the specific requirement, different forms of MS - including tandem MS (MS/MS) and top-down MS - are implemented for the definitive identification of conjugation points.
Peptide Mapping
This methodology involves the partitioning of the protein into smaller peptide subunits with the aid of proteases. The resultant peptides are subject to analysis using certain advanced techniques such as Liquid Chromatography (LC) in conjunction with MS or tandem MS. By correlating the gleaned peptide masses or fragmentation patterns with the theoretical sequence of the source protein, the conjugation positions can be distinctly established.
Edman Degradation
Edman degradation serves as a powerful instrument in sequential delineation of amino acids constituting a peptide or protein. By marking and severing the modified amino acid or the resultant conjugate from the protein, the altered residues can be conclusively identified.
X-ray Crystallography
X-ray Crystallography A potent strategy for discerning the three-dimensional configuration of proteins is X-ray crystallography. If a protein exhibits a conjugation or modification, it can be effectively visualized and ascertained in the electron density map derived from the gathered crystallographic data.
Progress of Conjugation Position Analysis
Conjugation Position Analysis Applications
Bioconjugation and antibody-drug conjugates (ADCs)
Conjugation position analysis is crucial in the development and production of ADCs, which involve attaching a drug molecule to an antibody. Knowing the exact site of conjugation helps ensure the efficacy and safety of the ADC.
Protein engineering
Determining the conjugation position is important in protein engineering to precisely modify or add functional components to proteins. This allows for the development of tailored proteins with improved properties or specific functionalities.
Drug delivery systems
Conjugation position analysis is utilized in the design and characterization of drug delivery systems, such as liposomes or nanoparticles. Identifying the conjugation site enables the controlled and targeted release of drugs to specific tissues or cells.
Diagnostic assays
Analyzing the conjugation position is important for the development and optimization of diagnostic assays, such as enzyme-linked immunosorbent assays (ELISAs) or lateral flow tests. Accurate positioning of conjugates ensures sensitive and specific detection of target analytes.
Want to Learn More?
Dedicated to assisting our clients in meeting the escalating and changing demand for dendritic polymer goods, Creative Proteomics is a rapidly expanding, preeminent global provider of research services and solutions. We offer protein drug characterization services for a variety of proteins such as protein structure confirmation, post-translational modification (PTM) analysis, protein physicochemical property determination, protein purity detection, protein impurities testing, and protein biosafety analysis. Please contact us if you have any questions about our services or are interested in collaborating with us. We would be happy to hear from you.
Reference
- Zhu, X., et al.; Current LC-MS-based strategies for characterization and quantification of antibody-drug conjugates. Journal of Pharmaceutical Analysis. 2020, 10(3), 209–220.
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For research use only, not intended for any clinical use.
