Page 47 - Pharmaceutical- Guide to Biopharmaceutical

Characterization Quality Control Pharmacokinetics

Pretreatment for Quantitative Analysis of Antibodies nSMOL Antibody BA Kit

Pretreatment Kit for LC/MS/MS Quantitative

Pretreatment Method Developed for Analysis of Monoclonal Antibodies

Quantitative Analysis of Monoclonal Cell Line Optimization

Antibodies in Blood by LC-MS/MS click here
benefits

Operating Principle and Features of Kit Resin surface • Selectively recovers Fab peptides and enables analysis without excessive peptides or trypsin.
Immunoglobulin Culture
The nSMOL Antibody BA kit can be used for all types of antibody Collection • Offers general applicability for any type of antibody drug.
Antibodies
drugs and eliminates the need for creating and cross-testing antibodies • No antibodies or ligands are needed for capturing, which streamlines method
specifically for detection, which is typically required for detecting
monoclonal antibodies. Advantages include a broader dynamic range 5 μL Plasma Immunoglobulin collection resin development and helps reduce costs.
and much higher selectivity than the ELISA method and the ability to Add Binding solution
analyze multiple components simultaneously. In terms of operation,
performing the pretreatment on filter cups avoids tedious washing
operations and samples can be collected easily by centrifuging (Fig. 1). Capturing
After pretreatment, samples can be injected directly into the LC-MS
system. On the plastic surface, the antibody collection resin is coated Centrifuge Wash: Wash solution 1
to ensure the Fab fragment side of antibodies is oriented toward the Purification
outside. By adding nanoparticles with trypsin solidified on the surface Centrifuge Wash: Wash solution 2
to the antibodies, the trypsin can access the antibodies efficiently for
trypsin digestion (Fig. 2). The peptides derived from Fab fragments by Add Reaction solution + ISTD Nano particle
trypsin digestion can be collected easily by centrifuging. (FG beads Trypsin DART)
Selective digestion
of Fab region Trypsin-accessible
surface
Centrifuge at 50 ºC for
Sample collection 4 to 6 hrs
Fab peptides Characterization

MRM measurement Approx. 5 minute
of signature peptides LCMS analysis
Fig. 1 Simple Workflow

Trypsin-immobilized
nanoparticle Quality Control

Diameter: LCMS Analysis
200 nm
Kit Contents
Collection of Fab-derived
Captured Antibody Fab Limited Reagent Name Quantity Capacity Storage Temperature
Oriented toward Solution Trypsin Access peptide
Immunoglobulin collection resin 2 1.3 mL per vial 4 °C
Fab Wash solution 1 (Binding solution) 1 80 mL 4 °C
• Decreases sample Wash solution 2 1 80 mL 4 °C Pharmacokinetics
Antibody complexity Reaction solution 1 10 mL 4 °C
• Prevents contamination
IgG collection from excess proteolysis Enhanced reaction solution 1 Freeze-dried 4 °C
Reaction stop solution 1 1 mL 4 °C
Diameter: 100 nm FG beads Trypsin DART 1 1.1 mL -20 °C *1
Note: The reagent kit is shipped refrigerated (2 to 8 °C).
*1 For long-term storage of 1 month or more, store at -80 °C. Others
Immunoglobulin collection resin
Fig. 2 Operating Principle of Kit

46 47
index index