nSMOL Antibody BA Kit
LC/MS/MS Sample Prep Kit for Quantitative Analysis of Monoclonal Antibodies
nSMOL – How Nano Technology and LC-MS Improved Speed and Accuracy of mAb Bioanalysis
nSMOL (nano-surface and molecular orientation limited proteolysis) is Shimadzu’s proprietary, innovative technique that enables selective proteolysis of the Fab region of monoclonal antibodies. The nSMOL Antibody BA Kit is a ready-to-use reagent kit for collecting monoclonal antibodies from blood or other biological samples using immunoglobulin collection resin, and then performing selective proteolysis of the Fab region of these antibodies via trypsin-immobilized nanoparticles. Variable region-derived peptides produced by limited proteolysis can then be quantified via MRM measurements utilizing a high-performance LCMS-8050/8060 triple quadrupole liquid chromatograph mass spectrometer. An unparalleled convenient and rapid workflow provided by the nSMOL Antibody BA Kit dramatically improves the productivity and robustness of LCMS mAb bioanalysis.
- Faster, Less Expensive Method Development
Dramatically improves response and quantitative repeatability. No capture antibodies or ligands required.
- nSMOL Proteolysis
Selective collection of Fab peptides. Limits contamination from excessive peptides or trypsin.
Highly sensitive and accurate assays are possible for a variety of antibodies.
- Highly Versatile
Applicable to a wide variety of pharmaceutical antibodies.
This kit enables highly reproducible data and avoids the troublesome steps of denaturing, reduction, and alkyation normally associated with protein digestion. There is also no need for solid phase extraction after reaction. After nSMOL preparation, samples can be injected directly onto the LCMS.
The nSMOL Antibody BA Kit provides prepared reagents and protocols for sample prep via the nSMOL method.
|Detection Targets||Monoclonal antibodies in serum or plasma (including antibody pharmaceuticals)|
|Number of Measurement||100 assays|
|Approximate volume of sample||90 μl (before LCMS injection)|
|Storage Temperature||Specified for each reagent|
|Expiration Date||18 months from the date of shipment if unopened and stored at 4°C
(*For FG BeadsTM Trypsin DARTTM, 18 months from the date of shipment if stored at -80°C)
|Immunoglobulin Collection Resin||2||1.3 mL/each||4°C|
|Wash（Binding）Solution 1||1||80 ml||4°C|
|Wash Solution 2||1||80 ml||4°C|
|Reaction Solution||1||10 ml||4°C|
|Enhanced Reaction Solution||1||Freeze-dried||4°C|
|Reaction Stop Solution||1||1 ml||4°C|
|FG Beads™ Trypsin DART™||1||1.1 ml||-20°C*1|
*1 If it is being stored for a month or longer, store it at -80 °C.
The following consumables are purchased separately.
|nSMOL reaction socket tube||SHIMADZU||225-32260-91
|Ultrafree-MC, GV 0.22 μm||Merck Millipore||UFC30GV00
|Micro tube 2mL, PP||SARSTEDT||72.708|
Note: For other equipment required, please refer to instruction manual.
Refer to the recommended specifications and reference products in the following table, and make preparations accordingly.
|Required Items||Recommended Specifications||Reference Products|
|5 μm filtration filter||Prepare a syringe and syringe filter with a pore size of 5 μm.
Millex-SV 5.0 μm, PVDF, 25 mm,
|PVC housing, non-sterile,
SLSV025NB, Merck Millipore
|0.8 μm filtration filter||Prepare a syringe and syringe filter with a pore size of 0.8 μm.||Millex-AA Syringe Filter Unit, 0.80 μm,
mixed cellulose esters, 25 mm, PVC housing non-sterile,
SLAA025NB, Merck Millipore
|2 mL Sample Tube||・Polypropylene
・Round bottom tube
|Item name:Safe-Lock Tubes
|Micro Tube Mixer||・Used at room temperature or 50°C.
・Can be installed inside the incubator.
・Set available with a 1.5 mL tube, a 2.0 mL tube, and a filter tube.
|Item name:Micro tube mixer
Sold by:Tomy Seiko Co., Ltd
|Incubator||・Incubator with a humidistat, or an incubator that a tray filled with water can be placed inside in order to maintain saturation vapor pressure
・Big enough that the above-mentioned micro tube mixer can be installed
|Item name:Incubator with a freezer
Sold by:Panasonic Healthcare Co., Ltd.
|Micro Pipetter, Tip||Tip without plasticizer to avoid plasticizer contamination in the MS (colorless)||(1) Brand name:Rainin Sold by:Mettler Toledo
(2) Brand name:Biohit Sold by:Sartorius
|Desktop Centrifuge||Capable of centrifugation of 1.5 mL tubes, 2.0 mL tubes, and filter tubes at 10,000 μg and 12,000 μg||Product series name:MX series high speed micro centrifuge
Sold by:Tomy Seiko Co., Ltd
|Magnetic Tube Stand||Capable of magnetically separating FG beads trypsin DART||Item name:Magnet stand
Sold by:Tamagawa Seiki Co., Ltd.
|LCMS Vials||Low adsorption type, or vials without plasticizer Item name:TORAST-H Bio Vial P/N:GLCTV-H-BIO||Item name:TORAST-H Bio Vial P/N:GLCTV-H-BIO
Sold by:Shimadzu GLC
|Analysis Column||Capable of peptide separation and analysis||
Item name:Shim-pack GISS
|Internal Standard Solution||Capable of sensitivity and retention time correction||Item name:ProteoMass P14R MALDI-MS standard