Intact Mass Validation
To ensure the integrity and accuracy of the obtained antibody sequences, Creative Biolabs performs intact mass validation, ensuring the acquisition of functional antibodies in one go and establishing a reliable foundation for subsequent antibody engineering and therapeutic development.
Introduction of Intact Mass Validation
Intact mass validation involves using high-precision mass spectrometry that combines high ion transmission for large molecules to measure the masses of the heavy and light chains of antibodies. These measured values are then compared with the theoretical masses (expected masses calculated based on the antibody's sequence information) to ensure that no amino acid deletions or mutations are overlooked.
Importance of Intact Mass Validation
Ensuring Sequence Integrity and Accuracy
Intact mass validation is a critical step in ensuring the precise and accurate antibody sequence. By accurately measuring the masses of the heavy and light chains and comparing them with theoretical masses, potential amino acid deletions or mutations can be identified.
Focus on Heavy Chain Variability
The highly variable CDR3 region of the heavy chain necessitates stringent sequence integrity requirements. Intact mass validation ensures that these variable regions are not omitted or altered.
Early Detection of Issues
Intact mass validation enables the early detection of potential sequence issues, providing a reliable sequence foundation for subsequent antibody engineering, production, and therapeutic development, thus avoiding potential issues that could affect future applications.
Support for Functionality and Reliability
By ensuring the accuracy and integrity of the antibody sequence, intact mass validation guarantees the functionality and reliability of antibodies for applications such as antibody engineering, production, and therapeutic development.
Intact Mass Validation Process at Creative Biolabs
1. Reduction
Antibodies are first reduced using the reducing agent dithiothreitol (DTT) to break open the disulfide bonds between the light and heavy chains.
2. Heavy-Light Chain Separation
Reverse-phase high-performance liquid chromatography is used to separate the light and heavy chains.
3. Mass Measurement
The separated light and heavy chains are introduced into the high-precision mass spectrometry instrument, known for its high ion transmission efficiency and mass accuracy.
4. Theoretical Mass Comparison
The measured masses of the light and heavy chains are compared with theoretical masses.
5. Quality Difference Analysis
Any differences between the measured and theoretical masses can be identified through comparison.
Creative Biolabs Service Advantages
Creative Biolabs incorporates intact mass validation steps in every antibody sequencing service and other analysis service provided to clients, ensuring that the acquired antibody sequences are precise and complete, thereby forming a solid foundation for subsequent antibody application development. Contact us now to embark on a journey of excellence and reliability in antibody research and development!
Related Services:
- DASS Technology
- Leucine/Isoleucine Discrimination
- Isobaric Amino Acids Discrimination
- Manual Sequence Curation
- State-of-the-Art Instruments