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h The peak area of the non-CF glycopeptide (TEPPLnATAGDQEEK-N6H3; Uniprot ID: G3I973) from hypoxia upregulated protein 1 at different time points. Core fucosylation was evidenced by the ion consisting of the peptide and the glycan moiety (HexNAc(1)dHex(1)). c carbomidomethylation of ...
a protein sample in PBS requires a 91-fold dilution to achieve 50% of the potential MS signal. Therefore, if the protein concentration is greater than 90 µM and salt adducts will not impede data analysis, the sample can be diluted following Protocol 1. Otherwise, sample cleanup by ultr...
The flow rate was 1 mL/min and protein elution was determined by UV absorbance at 280 nm (Beckman System Gold HPLC with a model 166 or 167 UV detector, Beckman Instruments). Electrophoretic (Western) transfer and immunoblouing. SDS- PAGGE was performed under denaturing conditions except that...
Cation exchange-HPLC and mass spectrometry reveal C-terminal amidation of an IgG1 heavy chain. A unique, late-eluting "basic peak" (relative to the "main peak") was observed by weak cation exchange–HPLC (WCX) for a recombinant monoclonal antibody (m... KA Johnson,K Paisley-Flango,BS Tan...
11.An antibody expression plasmid, wherein a nucleotide sequence comprising the heavy chain variable region according to claim 1 and a mouse IgG1 heavy chain constant region expresses a heavy chain protein of a broad-spectrum intact recombinant antibody against clothianidin and dinotefuran. ...
quickly verify recombinant protein expression or monitor sample micro-heterogeneity. BGI scientists at the San Jose Mass Spec Center have developed cutting edge Intact Mass services including Native MS utilizing multiple HPLC separation strategies combined with high resolution Orbitrap mass spectrometry1,2,...
Since protein A and protein G show affinity towards the constant part of human IgG, carriers can be coated with one or a combination of both proteins in order to immobilize capturing antibodies or to extract therapeutic mAbs. There are positive and negative aspects of this kind of sample prepar...
The six most-abundant plasma proteins (albumin, IgG, IgA, transferrin, haptoglobin, and α-1-antitrypsin) consisting roughly of 90% of the total protein in the pooled aliquots were removed by immunodepletion chromatography (Multiple Affinity Removal Column HU-6, 4.6-mm inner diameter (ID) × ...
To determine the protein concentration, protein assays were performed as previously described [16]. For metabolite extraction, 700 µL ice-cold methanol (100%, HPLC grade) was added to freeze-dried cells and vortexed for 30 s. The mixture was sonicated at 4 °C for 30 min and then centr...