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Veterinary Therapeutic Antibody Characterization
The characterization of veterinary therapeutic antibodies is used to assess and validate the properties and quality of the produced monoclonal antibodies. BioVenic provides veterinary therapeutic antibody characterization services including both structural and functional characterization. These characterization steps help ensure the consistency, safety, and efficacy of the antibodies, meeting the requirements for subsequent therapeutic use in animals.
Fig.1 Monoclonal Antibody Structure. 1
Structural Characterization
- Primary Structure
The primary structure of veterinary therapeutic antibodies refers to their amino acid sequences, including both the light and heavy chains. It is the foundation for their specificity, affinity, functionality, classification, and antigen recognition. BioVenic utilizes LC/MS, which stands for liquid chromatography-mass spectrometry, to provide you with antibody primary structure characterization services. We determine the molecular mass of samples and analyze their amino acid sequences.
- Higher-Order Structures
The secondary structure of veterinary therapeutic antibodies includes elements such as α-helices, β-sheets, and random coils within the protein. It can impact the antibody's spatial conformation, thus affecting its binding to antigens. BioVenic has established a secondary structure characterization platform, utilizing techniques like circular dichroism spectroscopy (CD) and Fourier transform infrared spectroscopy (FTIR).
Circular Dichroism Spectroscopy
CD involves the differential absorption of left-handed circularly polarized light and right-handed circularly polarized light. CD spectroscopy can be conducted in solution, providing insights that are closer to the protein's physiological state. Moreover, this method is fast, straightforward, and highly sensitive to conformational changes.
Fourier Transform Infrared Spectroscopy
In FTIR, specific wavelengths of infrared light are directed onto a sample, where they can be either reflected or absorbed, and their interactions are measured. Different structural regions of proteins generate distinctive absorption bands, and this information can be analyzed to determine the veterinary therapeutic antibodies' secondary structure.
The tertiary structure refers to the spatial arrangement and interactions between amino acid residues in therapeutic antibody molecules, determining their specificity, affinity, and biological activity. BioVenic offers services for tertiary structure characterization, typically utilizing techniques such as X-ray crystallography and high-sensitivity spectroscopic methods like hydrogen-deuterium exchange mass spectrometry (HDX-MS).
X-Ray Crystallography
Obtaining the 3D structure of a protein is achieved through X-ray diffraction of the protein crystals. When X-rays are directed at the protein crystal, it scatters the X-rays onto an electron detector or photographic film. By rotating the crystal, 3D diffraction data is captured, allowing the calculation of the positions of each atom within the crystalline molecule.
Hydrogen-Deuterium Exchange Mass Spectrometry
When samples are dissolved in a liquid, they undergo an exchange with hydrogen atoms present in the solution. The rate of this exchange between hydrogen and deuterium provides the accessibility of solvent molecules within the protein structure and helps us infer the protein's structure and conformation.
- Post-Translational Modification
Disulfide bonds and glycosylation are typically associated with post-translational modifications of proteins, playing a crucial role in the spatial folding and stability of proteins. BioVenic provides mass spectrometry methods like collision-induced dissociation (CID) and high-energy collision dissociation (HCD), used for disulfide bond and polysaccharide composition information analysis.
Functional Characterization
- Antigen-antibody Interaction
Veterinary therapeutic antibody samples must specifically bind with the target antigen with high affinity to form antigen-antibody complexes in order to exert their intended effects within the treated animals. BioVenic provides functional characterization services of veterinary therapeutic antibodies, including the detection of antigen-antibody interaction, such as at the protein level using ELISA and SPR-based assays, as well as at the cellular level using fluorescence-activated cell sorting (FACS).
Surface Plasmon Resonance
This is a physical phenomenon based on optical, label-free detection technology, which, without the need for labeling or purification of various biomolecules, can be used for real-time detection of binding interactions between two or more molecules when incident light at a metal and dielectric interface satisfies certain energy and momentum matching conditions, thereby exciting the coherent oscillations of free electrons on the metal surface to generate surface plasmons.
- Cellular Function
BioVenic provides services that allow for cellular response detection and generated biomarkers following the antigen-antibody interaction. This aids in understanding the impact of the structure and chemical modifications of therapeutic antibodies in the veterinary field on their biological activity.
| Services | Description | Assays |
|---|---|---|
| Cell Proliferation Assays | They are used to monitor the response and health of cultured cells following antigen-antibody binding. |
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| Cell Apoptosis Assays | Programmed cell death, antibodies may induce or inhibit apoptosis depending on their application and mechanism. |
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| Antibody Endocytosis Assays | Influences the biological activity and homing of the antibody within the cell, helping to determine whether the antibody can be endocytosed by the cell. |
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| Cytokine Assays | Plays an important role in immunomodulation. |
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Why Choose Us?
Comprehensive characterization services from structure to function.
Selecting the appropriate characterization targets according to specific needs.
We optimize parameters and procedures to provide with high-quality characterization results.
BioVenic offers comprehensive characterization services and has established advanced technological platforms for specific needs. We thoroughly characterize the veterinary therapeutic antibody samples to ensure they meet your expectations in terms of results and functionality, meeting the therapeutic effects expected from veterinary antibodies. Don't hesitate to contact us if you require these services!
Reference
- Moorthy, Balakrishnan S., et al. "Structure of monoclonal antibodies." Biobetters: Protein Engineering to Approach the Curative (2015): 81-89.