Introduction
Immobilized antibody systems are the key to develop efficient diagnostics and separations tools. In immunoassays
(IAs), immobilized antibodies are preferred over their free counterpart to curtail the number of detection steps and
time, as well as to improve high detection sensitivity and reproducibility. Ideally, the immobilized antibody, an
antibody with immobilized material formed by linking the antibody to the surface of the stationary phase, should
maintain its natural conformation, and there is no space for antigen binding between adjacent antibody molecules
with the Fc fragment facing the surface of the solid phase, and the Fab fragment exposed to the liquid phase.
However, the binding of antibodies on the carrier may have different spatial curves, and some immobilized antibodies
may inactivate or denature due to random orientations that caused by space effects during immobilization, thereb
losing antigen-binding activity. Therefor, sequencing of immobilized antibodies is particularly important in order
to detect whether changes in protein conformation are caused during the immobilization of antibodies.
Fig.
1. Schematic diagram of IgG immobilization on the gold surface modified with a solution of CS2 and protein A. (Niu
Y, et al., 2012)
Our Services
As an advanced protein sequencing service provider, Creative Proteomics has a powerful protein
sequencing platform to accelerate your project's R&D journey. Here, we provide you with comprehensive
immobilized antibody sequencing services.
At Creative Proteomics, our highly qualified staff develops mass spectrometry platform-based de novo
sequencing services for your immobilized antibodies, and high-resolution mass spectrometers and advanced
bioinformatics analysis software guarantee a 100% success rate. You only need to provide 1mg of antibody (purity
>90%) for detection.
The following is our simple sequencing workflow:
Ⅰ. Immobilized antibody are digested with enzyme to obtain short overlapping peptides.
Ⅱ. LC-MS/MS of each peptide are achived by mass spectrometer.
Ⅲ. Based on bioinformatics software, data analysising including de novo peptide identification and automated
overlapping peptide assembly.
Ⅳ. Delivery of a comprehensive immobilized antibody sequencing report.
With our extensive sequencing experience, you can also expect:
- Customized service: Customized solutions for you according to your testing purpose and budget.
- High quality report: All raw data and processing are contained .
- Professional technology platform: Based on high resolution (greater than 105) and mass accuracy (less than 1ppm)
Mass spectrometry platform to ensure the accuracy of identification results.
- Advanced bioinformatics analysis: Protein sequences can be accurately identified without losing any valid data
information.
- Wide range of application: All antibodies are applicable theoretically.
Creative Proteomics has accumulated extensive experience in the discovery and sequencing of
therapeutic antibodies and proteins. We provide professional immobilized antibody sequencing services for global
customers. Our team has successfully sequenced thousands of antibodies to help researchers decode their antibodies
in a fraction of the time. Therefore, please contact us if you are
interested in our antibody sequencing services.
References
- Niu Y, Matos A I, Abrantes L M, et al. (2012). Antibody oriented immobilization on gold using the
reaction between carbon disulfide and amine groups and its application in immunosensing. Langmuir.
28(51):17718-25.
- Shen M, Rusling J, Dixit C K. (2016) Site-selective orientated immobilization of antibodies and conjugates for
immunodiagnostics development. Methods. 116:95-111.
