Protein Sequencing Solutions

The End-to-End
Solution for
Protein Insights

Next-gen protein sequencing (NGPS) delivers single-molecule precision with expanding coverage enabling targeted protein analysis at your benchtop.

Talk to an Expert

Protein Sequencing Kits

Achieve deeper protein insights with ready-to-sequence chips, recognizers, and aminopeptidases.
 

Protein Sequencing Kits

Library Preparation Kits

High-confidence protein sequencing from Nanogram inputs with Library Preparation Kit V3.
 

Library Prep Kits

Products Sequencing Solutions

Unlock Proteome Depth with Sequencing Kit V4

Accessible NGPS Icon

Accessible NGPS
Single-molecule resolution to identify and characterize proteins on a benchtop instrument

Insights Beyond ProlineIcon

Insights Beyond Proline
Navigate challenges posed by proline-rich domains, common in antibodies and membrane proteins, with Sequencing Kit V4

Expanded Coverage Icon

Expanded Coverage
New recognizers, peptidases, and improved bioinformatics offer an 85% increase in sequenceable peptides

Protein Mixture Precision Icon

Protein Mixture Precision
Enhanced bioinformatics to identify peptides with greater confidence

Sequencing Through Proline

Products Sequencing Solutions

  • V4 detects amino acids beyond proline, which accounts for 6% of the total amino acids in the human proteome and over 40% of peptides generated by Lys-C digestion
  • Proline is abundant in vital proteins that are difficult to analyze by mass spec, common in membrane proteins, antibodies, and transcription factors

Proline’s unique structure impacts the peptide fragmentation for mass spec analysis, a phenomenon referred to as the “proline effect,” which restricts sequencing by mass spec. The V4 kit delivers improved chemistry that sequences through proline rich regions, expanding sequencing insights on Platinum.

Expanded Proteome Coverage

Upgraded chemistry improves single-molecule sequencing by adding glycine (G) and methionine (M) recognition and enhancing alanine (A) and serine (S) recognition. The inclusion of X-prolyl aminopeptidase allows for cleavage of proline-rich peptides. Improved bioinformatics, including enhancements to amino acid calling and alignment algorithms, delivers higher-quality data with V4, enabling richer data capture on the sequencer.

Expanded coverage achieved by the V4 Sequencing kit was demonstrated with panel of ten recombinant proteins that were digested into peptides, and sequenced using the Platinum Pro with both the V4 and V3 kits showing:

  • Increased peptides coverage in 9 of 10 proteins
     
  • Increased peptide alignments (sequenced copies) in 10 of 10 proteins
     
Chart: Mean (Peptides Identified) and Mean (Alignments) VS Sample
Sequencing ten recombinant proteins, from 15 to 66 kDa, demonstrates that the V4 kit produces more peptides and alignments than the V3 kit, with several peptides detected only by V4. V3 sequencing data was analyzed with analysis software version 2.15, and V4 was analyzed with version 2.18 optimized for recognizers provided in the V4 sequencing kit.

Protein Mixture Precision

Upgrades to the analysis software boost confidence in accurate protein identification in mixtures using V4. An equimolar mixture of ten proteins sequenced with V4 demonstrated increased peptide alignments, resulting in increased precision in peptides identified compared to V3.

 

Chart: plot graph illustrating the improvement of protein id confidence when comparing an equimolar mix of 20 proteins of different sizes vs previous kit
Upgrades to the analysis software boost confidence in accurate protein identification in mixtures using V4 when comparing an equimolar mix of 10 proteins of different sizes. V3 sequencing data was analyzed with analysis software version 2.15, and V4 was analyzed with version 2.18 optimized for recognizers provided in the V4 sequencing kit.

Sequence Antibodies

Prolines play a crucial role in antibody structure and function, shaping complementarity-determining regions (CDRs) and immunoglobulin loops. We sequenced trastuzumab, an anti-HER2 monoclonal antibody used in cancer treatment, on the Platinum Pro, to demonstrate the improved coverage obtainable with V4, exhibiting:

  • 2.5-fold more peptide coverage in heavy and light chains compared to V3
     
  • Significant increase in amino acid calls after proline with V4 chemistry

 

Chart: Sequencing of light and heavy chains of trastuzumab with Sequencing Kit V4

Sequencing of light and heavy chains of trastuzumab with V4 and Binder Order sequencing workflow v2.18 demonstrates a significant increase in amino acid coverage obtained through glycine (G) and methionine (M) recognition, enhanced alanine (A) and serine (S) recognition, and identification of amino acids beyond proline (P) compared to Sequencing Kit V3 analyzed with software v2.15. LC: light chain; HC: heavy chain.

Characterize Proteins by Immunoprecipitation

Traditional methods for identification of proteins enriched by immunoprecipitation (IP) vary in their resolution, where Western blot offers qualitative to semi-quantitative measurements that are antibody-dependent, and mass spectrometry can produce exhaustive data inclusive of proteins that may be nonessential to a study. Platinum Pro with the V4 kit is an accessible non-antibody-dependent, higher-resolution option optimized to identify target proteins with concise data readouts.

  • Sequence IP products on Platinum Pro for clear identification of protein interactors
     
  • Explore signaling pathways, antibody binding partners, and other protein interactions

 

Chart: Heat shock protein (HSP90) enriched from MCF-7 cells and sequenced with V4
Heat shock protein (HSP90) enriched from MCF-7 cells, sequenced with V4, and analyzed with Binder Order sequencing workflow v2.18 correctly identified HPS90 A and B as top-ranked inference with an FDR <10% using the whole human proteome. C: crude; FT: flow-through; W: wash; B: beads.

Explore our resources to discover how next-gen protein sequencing with Platinum Pro can enhance your protein research

Have questions?