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There's a massive leap forward into the new era of this field, with unchallenged ability to achieve 100% accuracy in antibody sequencing by precisely distinguishing between leucine and isoleucine, based...
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There's a massive leap forward into the new era of this field, with unchallenged ability to achieve 100% accuracy in antibody sequencing by precisely distinguishing between leucine and isoleucine, based on the proprietary Database Assisted Shotgun Sequencing (DASS) technology.
show less
2 DEC 2019 · (1) The masses of the reduced light- and heavy-chain will be determined to validate the determined sequences. The light chain sequence must match within a +/- 1.2 Da range. For the heavy chain, +/- 1.8 Da is allowed. If the determined sequences do not match with the protein mass, you will not be charged. (Please note that this deviation is less than the weight of two protons).
(2) Each mutation in the antibody will be validated by at least two meaningful spectra. This ensures that your sequence does not contain isobaric exchanges. Do not rely on peptide maps alone. [Isobaric exchanges are exchanges of identical mass like GG=N, YM=FM (Ox). The peptide maps used by some competitors are often not sensitive to this kind of error]. http://www.creative-biolabs.com/next-generation-antibody-sequencing.html
2 DEC 2019 · Leucine can be distinguished from Isoleucine.
W can be distinguished from GE, AD and SV (by mass accuracy).
R can be distinguished from GV (by mass accuracy).
Q can be distinguished from K (by mass accuracy).
N can be distinguished from GG. Our DASS technology ensures bond breakage between GG. In addition, we use our proprietary derivatization technology for asparagine to ensure that it is not GG.
Q can be distinguished from GA. Our DASS technology ensures bond breakage between GA. In addition, we use our proprietary derivatization technology for glutamine to ensure that is not GG.
2 DEC 2019 · IgMs can usually be sequenced like normal IgGs. Slightly more sample may be required. As the constant region is modified by several glycans, which cannot be cleaved by PNGase F, we can only guarantee complete coverage for the variable part of the antibody.
Our method works with all antibody formats from all species (mouse, rat, hamster, llama, rabbit, etc.; Fab, scFv, etc.).
Some mAbs are coupled to matrices. This is compatible with our sequencing method, but more samples may be required.
Many older hybridomas may produce 2 light chains (one is the kappa-chain of myeloma MOPC 21). These mixtures can usually be sequenced, even when the MOPC 21 kappa chain is in two-fold molar excess.
We also provide DASS-Based Protein and Enzyme Sequencing service including full end to end sequencing, determination of post-translational modifications, intact mass, N-glycosylation sites, and artificial tags, etc.
2 DEC 2019 · Sequencing of Fluorochrome mAbs, IgMs, and other Non-Standard Antibodies
We have over 90% chance to get the sequence from tiny amount of your precious sample (e.g. 20 µg).
We can enrich even very dilute sample (e.g. western blot antibody) for sequencing.
Our method is usually not affected by ligands (FITC, Biotin, Alexa) coupled to antibodies. Larger protein ligands (PE, APC) make sequencing slightly more challenging. However, this can be compensated by using a higher protein concentration, because sequencing quality is dependent on sample amount.
2 DEC 2019 · Our method generates many overlapping peptides during the fragmentation process, enabling us to sequence very long stretches of unknown amino acids. The high quality of MS/MS spectra in combination with intelligent data mining, allows us to read the CDR3 like a book. The technique is so powerful that we were able to sequence a 20 kDa protein, which had no homolog in the database. http://www.creative-biolabs.com/next-generation-antibody-sequencing.html
2 DEC 2019 · De Novo Sequencing of the CDR3 Region
While the CDR3 of the light chain is mostly encoded by the germline sequences, the CDR3 of the heavy chain is usually not available in databases. It is encoded by the so-called D-segments, but these are modified by nucleases and terminal transferases. Typically, only 1-4 AA of a D-segment remains in the matured antibody. The rest of the D-segment is “artificial” and has to be sequenced de novo. http://www.creative-biolabs.com/next-generation-antibody-sequencing.html
2 DEC 2019 · Due to the high number of peptides, we get sequence information for EVERY peptide bond in the antibody. Typically, 20-70 different MS/MS spectra are generated for each amino acid (AA) position. Hence, even the hardest sequences of proline and arginine-rich peptides can be resolved. As the order of all amino acids is clear, there is no need for time-consuming techniques like Edman sequencing. http://www.creative-biolabs.com/next-generation-antibody-sequencing.html
2 DEC 2019 · One-Stop De Novo Antibody Sequencing Services
Sequencing of the V and J and C Segments by DASS
The V and J and C gene segments of antibodies are available in public databases. However, during the maturation of an antibody, the B cell introduces hypermutations into the sequence to optimize the affinity. Our mapping algorithm is error tolerant and can match the "mutated" peptides to the corresponding germline, reliably.
There's a massive leap forward into the new era of this field, with unchallenged ability to achieve 100% accuracy in antibody sequencing by precisely distinguishing between leucine and isoleucine, based...
show more
There's a massive leap forward into the new era of this field, with unchallenged ability to achieve 100% accuracy in antibody sequencing by precisely distinguishing between leucine and isoleucine, based on the proprietary Database Assisted Shotgun Sequencing (DASS) technology.
show less
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| Author | echo han |
| Organization | echo han |
| Categories | Health & Fitness |
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