Nanoacquity chromatographic system

Manufactured by Waters Corporation

Sourced in United Kingdom, Germany

The NanoAcquity chromatographic system is a high-performance liquid chromatography (HPLC) instrument designed for the separation and analysis of small-volume samples. It is capable of handling flow rates from 20 nL/min to 2 μL/min, making it suitable for applications that require high sensitivity and low sample consumption.

Automatically generated - may contain errors

Lab products found in correlation

Compact qtof mass spectrometer, by Bruker (2 mentions) Pepmap 100 c18 trap column, by Thermo Fisher Scientific (1 mentions) Compact mass spectrometer, by Bruker (1 mentions) Tripletof 5600 instrument, by AB Sciex (1 mentions) Spectronaut software, by Biognosys (1 mentions)

Spelling variants of this product

NanoAcquity (157 citations) NanoAcquity system (49 citations) Chromatographic system (7 citations) NanoACQUITY UPLC® chromatographic system (2 citations)

5 protocols using nanoacquity chromatographic system

Glycopeptide Separation and Profiling by Nano-LC-MS/MS

Check if the same lab product or an alternative is used in the 5 most similar protocols

Digested glycopeptides were separated on nanoAcquity chromatographic system (Waters, Milford, MA) coupled to Compact mass spectrometer (Bruker, Bremen, Germany) with an electrospray ionization (ESI) source. Samples were loaded either directly after the overnight trypsin digestion (2 μL from 20 μL) or after the enrichment procedure (20 μL). They were loaded onto a PepMap 100 C18 trap column (5 mm x 300 μm, Thermo Fisher Scientific) at a flow rate of 40 μL/min of solvent A (0.1% formic acid) to wash off impurities and salts. Glycopeptides were separated on C18 analytical column (150 mm x 100 μm, 100 Å, Advanced Materials Technology) using a linear gradient from 0% to 80% of solvent B (80% ACN) in solvent A, at a flow rate of 1 μL/min in a 90-minute analytical run.
Fragmentation of glycopeptides was performed by tandem MS/MS by using CaptiveSpray interface, where nanoBooster was used to introduce gaseous acetonitrile into nitrogen flow. The mass spectrometer operated in positive ion mode; capillary voltage was set to 1300 V, nitrogen pressure was set to 0.2 bar, and the drying gas to 4.0 l/min at 150°C. Auto MS/MS method was used by selecting three precursor ions and exclusion criteria after three MS/MS spectra. Mass range was from 50 m/z to 4000 m/z, with a spectra rate of 1 Hz. Transfer time was from 70 μs to 150 μs, and pre-pulse storage was 12 μs.

Nakić N., Tran T.H., Novokmet M., Andreoletti O., Lauc G, & Legname G. (2021). Site-specific analysis of N-glycans from different sheep prion strains. PLoS Pathogens, 17(2), e1009232.

+ Open protocol

+ Expand

Quantitative Yeast Proteome Profiling

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

For protein profiling, single colonies of respective yeast strains were grown to mid-log phase (OD₆₀₀ ~0.7) in selective medium containing 2 % glucose, harvested by centrifugation and snap-frozen in aliquots equalling 10 OD₆₀₀ units. Sample preparation was carried out as previously described ^{60 (link)}. SWATH LC-MS/MS analysis was performed essentially as previously described ^{60 (link)} on a TripleTOF5600 instrument (SCIEX) online coupled to a nanoACQUITY chromatographic system (Waters) operating at 3 μL/min flow rate. Data was analyzed with Spectronaut software (version 14, Biognosys AG) and post-processed in statistical language R. Principal component analysis was carried out using the unfiltered data set and a prcomp function. Protein fold change was calculated with reference to the wild type strain, and differential abundance was defined as a fold change > 1.5 and FDR-corrected p-value < 0.01. Pathway enrichment was performed in String-db v11 ^{111 (link)}, and Reactome pathways ^{112 (link)} were reported as significantly enriched if FDR-corrected enrichment p-value < 0.05.

Vowinckel J., Hartl J., Marx H., Kerick M., Runggatscher K., Keller M.A., Mülleder M., Day J., Weber M., Rinnerthaler M., Yu J.S., Aulakh S., Lehmann A., Mattanovich D., Timmermann B., Zhang N., Dunn C.D., MacRae J.I., Breitenbach M, & Ralser M. (2021). The metabolic growth limitations of petite cells lacking the mitochondrial genome. Nature metabolism, 3(11), 1521-1535.

+ Open protocol

+ Expand

IgG N-Glycopeptide Analysis Protocol

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Sample preparation and analysis of IgG N-glycopeptides was done following a previously described protocol with minor changes [34 (link)]. Briefly, IgG was isolated from 90 µL of serum samples by affinity chromatography using CIM^® 96-well Protein G monolithic plate (BIA Separations, Ajdovščina, Slovenia). IgG N-glycopeptides were prepared by trypsin digestion of an aliquot of IgG isolates (25 μg on average per sample) followed by reverse-phase solid phase extraction (RP-SPE). Purified tryptic IgG N-glycopeptides were separated and measured on nanoAcquity chromatographic system (Waters, Wilmslow, UK) coupled to Compact Q-TOF mass spectrometer (Bruker, Bremen, Germany), equipped with Apollo II source and operated under HyStar software version 3.2. The first four isotopic peaks of doubly and triply charged signals, belonging to the same glycopeptide species, were summed together, resulting in 20 Fc N-glycopeptides per IgG subclass. Predominant allotype variant of IgG3 tryptic peptide carrying N-glycans in the Caucasian population has the same amino acid sequence as IgG2 [35 (link)]. Therefore, IgG glycopeptides were separated into three chromatographic peaks designated as IgG1, IgG2/3 and IgG4. Signals of interest were normalised to the total area of each IgG subclass.

Monaghan T.M., Biswas R.N., Nashine R.R., Joshi S.S., Mullish B.H., Seekatz A.M., Blanco J.M., McDonald J.A., Marchesi J.R., Yau T.O., Christodoulou N., Hatziapostolou M., Pucic-Bakovic M., Vuckovic F., Klicek F., Lauc G., Xue N., Dottorini T., Ambalkar S., Satav A., Polytarchou C., Acharjee A, & Kashyap R.S. (2021). Multiomics Profiling Reveals Signatures of Dysmetabolism in Urban Populations in Central India. Microorganisms, 9(7), 1485.

+ Open protocol

+ Expand

IgG N-Glycopeptide Profiling Protocol

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Sample preparation and analysis of IgG N-glycopeptides was done following a previously described protocol with minor changes [30 (link)]. Briefly, IgG was isolated from 90 µL of serum samples by affinity chromatography using a CIM 96-well Protein G monolithic plate (BIA Separations, Ajdovscina, Slovenia) and vacuum manifold. IgG N-glycopeptides were prepared by trypsin digestion of 25 µg of IgG isolates and purified with reverse-phase solid phase extraction (RP-SPE) using Chromabond C18 beads suspension applied to the wells of an OF1100 96-well polypropylene filter plate (Orochem Technologies Inc., Naperville, IL, USA) and vacuum manifold. Purified tryptic IgG N-glycopeptides were separated and measured on a nanoAcquity chromatographic system (Waters) coupled to a Compact Q-TOF mass spectrometer (Bruker, Bremen, Germany) equipped with an Apollo II source and operated under HyStar software version 3.2. After calibration, the first four isotopic peaks of doubly and triply charged signals belonging to the same glycopeptide species were extracted and summed together, resulting in 20 Fc N-glycopeptides per IgG subclass. Signals of interest were normalised to the total area of each IgG subclass.

Monaghan T.M., Duggal N.A., Rosati E., Griffin R., Hughes J., Roach B., Yang D.Y., Wang C., Wong K., Saxinger L., Pučić-Baković M., Vučković F., Klicek F., Lauc G., Tighe P., Mullish B.H., Blanco J.M., McDonald J.A., Marchesi J.R., Xue N., Dottorini T., Acharjee A., Franke A., Li Y., Wong G.K., Polytarchou C., Yau T.O., Christodoulou N., Hatziapostolou M., Wang M., Russell L.A, & Kao D.H. (2021). A Multi-Factorial Observational Study on Sequential Fecal Microbiota Transplant in Patients with Medically Refractory Clostridioides difficile Infection. Cells, 10(11), 3234.

+ Open protocol

+ Expand

Characterizing IgG1-Fc Proteins by ESI-MS

Check if the same lab product or an alternative is used in the 5 most similar protocols

IgG1-Fc proteins were analyzed by electrospray ionization mass spectrometry (ESI-MS). ESI spectra of reduced protein samples (15 mM dithiothreitol, pH 7.0, room temperature) were acquired on a SYNAPT G2 hybrid quadrupole / ion mobility / time of flight mass spectrometer (Waters Corp., Milford, MA). The instrument was operated in a sensitivity mode with all lenses optimized on the MH+ ion obtained from Enkephalin. The sample cone voltage was 30eV. Argon was admitted to the trap cell that was operated at 4eV for maximum transmission. Spectra were acquired at 9091 Hz pusher frequency covering the mass range from 100 to 3000 u and accumulating data for 1.5 seconds per cycle. Time to mass calibration was made with Nal cluster ions acquired under the same conditions. Samples were desalted on a reverse phase C4 column, 1 cm, 1 mm I.D. (Vydac, Midland, Canada, 300 A pore size. The 5 μm particles were packed by Micro-Tech Scientific) using a NanoAcquity chromatographic system (Waters Corp., Milford, MA). The solvents used were A (99.9% H₂O, 0.1% formic acid) and B (99.9% acetonitrile, 0.1% formic acid). A short gradient was developed from 1 to 70% B in 4 min with a flow rate of 20 μl/min. Masslynx 4.1 software was used to collect the data. The MaxEnt 1routine was used for processing data to convert peaks of multiply charged protein ions into uncharged deconvoluted protein spectra.

ALSENAIDY M.A., OKBAZGHI S.Z., KIM J.H., JOSHI S.B., MIDDAUGH C.R., TOLBERT T.J, & VOLKIN D.B. (2014). Physical stability comparisons of IgG1-Fc variants: effects of N-glycosylation site occupancy and Asp/Gln residues at site Asn 297. Journal of pharmaceutical sciences, 103(6), 1613-1627.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!