Peptide and Protein Bioanalysis: Infographic
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1. [ PEPTIDE AND PROTEIN BIOANALYSIS ] Aspirin 180 Da Bevacizumab 149 kDa Insulin Glargine 6,063 Da PEPTIDES AND PROTEINSSMALL MOLECULES Multiple precursor charge states dilute signal Increased fragmentation further reduces signal 2+ 3+ 4+ 5+ 1+ In 2013, 7 of the top 10 best-selling prescription drugs were large molecules 2 Humira 148 kDa Enbrel 150 kDa Remicade 149 kDa Lantus 6064 Da Rituxan 145 kDa Avastin 149 kDa Herceptin 148 kDa Seretide 500 Da Crestor 1001 Da Abilify 448 Da #1 #2 #3 #4 #5 #6 #7 #8 #9 #10 Increased sensitivity and specificity challenges: Fewer molecules in equal volume of analyte WHYis it harder to quantify large molecules? WHYare biotherapeutics more important than ever? HOW can Waters help? Optimize specificity of sample prep Oasis MAX and WCX SPE Elution Plates help achieve high recovery and low limits of detection for a wide range of endogenous, therapeutic and surrogate peptides. Wide mass range, reliable sensitivity The Xevo family of tandem quadrupole mass spectrometers delivers high sensitivity at high and low mass, detecting all fragments without compromise, while sensitivity at high m/z enhances specificity. Improved microfluidics for better sensitivity and reduced sample load The ionKey/MS System integrates the UPLC analytical separation directly into the source of the mass spectrometer, delivering significant increases in sensitivity, solvent savings, and reductions in sample volume. Increasing sensitivity through LC optimization $$ 10x UP TO IMPROVEMENT IN SENSITIVITY More LC and MS parameters to consider reduces efficiency Complex sample prep means possible loss of analyte STEP 1 STEP 2 STEP 3 STEP 4 Year 201420102005200019951990 204 9940 1.2.801.201.00 1.00 385 568 2.402.001.201.00 MRM of 1 Channel ES+ 1.32e4 Area 365 240 1.251.00 1.25 60 C 40 C 400 L/min 200 L/min ACN + 0.1% formic acid ACN + 0.1% formic acid + 5% TFE 14237 Temperature Flow rate Mobile phase B composition CAPILLARY VOLTAGE MOBILE PHASE COMPOSITION CHROMATOGRAPHIC PORE SIZE COLUMN TEMP FLOW RATE Analyzing large molecules may be one of the greatest challenges that the bioanalyst faces at the beginning of the 21st century. PEPTIDES AND PROTEINS ARE NOT SMALL MOLECULES. WHY TREAT THEM THE SAME? Biotherapeutics publications1 Use a generic set of chromatographic conditions and evaluate with the ACQUITY UPLC BEH C18 300 , ACQUITY UPLC CSH, and CORTECS UPLC C18 + Columns to identify the best peak and separation and then optimize key parameters, such as column temperature, flow rate, and mobile phase composition. Endogenous peptides and proteins yield high background SENSITIVITY SPECIFICITY 2015 Waters Corporation 720005295EN February 2015 AW-KW Learn more with the Peptide Bioanalysis Solution Guide Search literature code 720004563EN at waters.com 1 Electronic search performed in PubMed.org on January 13, 2015 with keyword biotherapeutics. 2 Kollewe, J. (2014, March 27). The world's 10 best-selling prescription drugs. The Guardian. Retrieved Jan. 13, 2015.