ENHANCED PROTEIN AND PEPTIDE CHARACTERIZATION

Transcription

1 Agilent MassHunter BioConfirm Software ENHANCED PROTEIN AND PEPTIDE CHARACTERIZATION Spectrum Algorithm Peak Modeling Deconvolution Mass Sequence Name Myoglobin (horse) Target mass Delta ppm 1.7 MinZ 6 MaxZ 18 Z Count 13 Fit Score U

2 AGILENT MASSHUNTER BIOCONFIRM SOFTWARE ENVISION HIGHER CONFIDENCE IN YOUR PROTEIN/PEPTIDE CHARACTERIZATION Agilent s sophisticated MassHunter BioConfirm Software brings you greater confidence in your protein/peptide analysis. Together with Agilent s high performance liquid chromatography/mass spectrometry (LC/MS) systems, it provides an intelligent solution for rapid confirmation of synthetic peptides and characterization of proteins and peptides. Now you can elevate your discovery by confirming your data with confidence and ease. BioConfirm Software Delivers: Precise and rapid protein confirmation Easier batch-to-batch analysis Comprehensive peptide mapping 2

3 ENVISION PRECISE PROTEIN CONFIRMATION BioConfirm provides both classical maximum entropy deconvolution and enhanced peak modeling (pmod) to determine the molecular weight of intact proteins. Intact Protein Workflow Qualitative Compound Report Data File Intact_MyoglobinNewMyo-112.d Sample Name NewMyo-1 Sample Type Sample Position P1-B-3 Instrument Name MH Instrument User Name wadmin Intact_MyoglobinNovartis.m Acq Method Success IRM Calibration Status Protein Sequence Mass Mod Profile1 Custom 3/4/214 2:8:13 PM BioConfirmIntactProteinHighMass-Default.m Column3 Column2 Info. Column3 horse_myo.txt Mod Profile Myo Protein Sequence 1 Myoglobin2.psq Mass Phospho&Ox A,B,C Custom1 test Stream Name S1 Walkup Method Description Intact Acq + Myo-DA Acquisition SW Version 62 series TOF/65 series Q-TOF B.6. (B52) Compound Table Mass Acquired Time DA Method IntProt Comment Column2 Sample Group Sequence Name Target Mass Myoglobin (horse) Delta ppm Height 1.7 MinZ MaxZ 6 Z Count 18 Fit Score 13 Uncertainty Significance.5 Algorithm 1 # Hits Peak Modeling Deconvolution Data File 3 Rule Intact_MyoglobinNewMyo-112.d Intact protein Spectrum Spectrum Algorithm Peak Modeling Deconvolution Acquire data Integrate and extract MS Match protein sequences and predict PTMs Deconvolute Mass MS Spectrum Peak List Calc m/z Diff(ppm) z Delta ppm 1.7 MinZ 6 MaxZ 18 Z Count 13 Fit Score Uncertainty.5 Significance 1 Rule Intact protein Abund Target mass Generate report MS Spectrum m/z Sequence Name Myoglobin (horse) End Of Report --- Maximum entropy deconvolution provides rapid mass resolution, especially for simple protein mixtures. For complex protein mixtures, the enhanced pmod algorithm removes artifacts from the spectrum and helps to resolve overlapping peaks even severely overlapping peaks providing cleaner spectra and more precise mass measurement. This enables fast confirmation of intact protein sequences and modifications. x x Printed at: 3:13 PM on:3/14/214 Page 1 of 1 (a) Maximum (b) Peak Modeling Counts vs. Deconvoluted Mass (amu) Deconvoluted spectra. (a) Maximum entropy deconvoluted spectrum. (b) Peak modeling deconvoluted spectrum. 3

4 ENVISION POWERFUL PEPTIDE MAPPING CAPABILITIES BioConfirm provides enhanced processing of MS/MS data for streamlined mapping and data interpretation. Peptide Mapping Workflow Acquire data Extract compounds Match Protein sequences and identify PTMs Visualize sequence coverage Generate report BioConfirm identifies peptides and PTMs based on peptide masses and product ions (b, y, and immonium ions) in the MS/MS spectra. This allows for faster and more streamlined processing of MS/MS data for peptide mapping. x , 1,1 Counts vs. Mass-to-Charge (m/z) 4 Peptide MS/MS spectrum product ion assignment.

5 ENVISION EASY BATCH-TO-BATCH COMPARISON BioConfirm provides easy visual comparison among samples, allowing for fast batch-to-batch analysis on both protein and peptide levels. Mirror Plot functionality enables rapid and reliable comparison of two samples, such as two batches of an engineered protein or biosimilars an essential part of biopharmaceutical analysis. Samples in the Mirror Plot can be switched quickly without reprocessing the data. Monitor Biosimilars and Batch-to-batch Variation by Quickly Comparing Samples and Reference The comparative analysis module facilitates the identification and visualization of the differences between two batches of data for easy inspection. Samples can be compared to a reference using chromatograms, MS, and MS/MS spectra. x Counts vs. Deconvoluted Mass (amu) Mirror Plot of two engineered proteins. x x ,2 1,35 1,5 1,65 Counts vs. Mass-to-Charge (m/z) x , 1, , 1,3 Counts vs. Mass-to-Charge (m/z) MS/MS Spectrum comparison results. MS Spectrum comparison results. 5

6 ENVISION RAPID AND ACCURATE mab GLYCAN ANALYSIS BioConfirm provides an easy and effective workflow for the analysis of mab glycans. mab Glycan Workflow N Acquire data Extract compounds Search database and identify glycans Calculate percentage and generate report BioConfirm enables users to accurately identify and visualize glycan structures in their mab samples using Agilent s Glycan Personal Compound Database (PCD) and extensive predefined glycan modifications. Using Agilent s mab-glyco Chip, customers have a comprehensive workflow for glycan analysis including hardware, software, and consumables a complete workflow that includes on-chip deglycosylation of the mab as well as enrichment, chromatographic separation, and detection of the cleaved glycans. All activities, including data processing using BioConfirm, can be completed in less than 15 minutes. 6

7 ENVISION WALK UP ANALYSIS FOR NON-EXPERT MS USERS BioConfirm part of Agilent s MassHunter Walkup solution. Sample Submission in Three Easy Steps Login Enter sample information Place vial in ALS Reports delivered to inbox MassHunter Walkup provides a simple-to-use interface that provides novice users the ability to analyze protein and peptide samples in three easy steps with minimal training. Users do not need to concern themselves with complex mass spec conditions; instead, they can run their samples with standard acquisition and data processing methods. Results are directly delivered to their Inbox. 7

8 Learn more Buy online Find a local Agilent customer center USA and Canada agilent_inquiries@agilent.com Europe info_agilent@agilent.com Asia Pacific inquiry_lsca@agilent.com Research use only. Information, descriptions, and specifications in this publication are subject to change without notice. Agilent Technologies shall not be liable for errors contained herein or for incidental or consequential damages in connection with the furnishing, performance, or use of this material. Agilent Technologies, Inc., 214 Published in the USA, April 7, EN