Home » Biologics CDMO » Bioanalytical » Protein/Ab Analytics

Key Features

  • Comprehensive breadth and innovative technologies
  • Versatile analytical procedure and strong capabiity in method development
Analytical services Analysis target Available method
Identification & characterization Protein identification Western Blot
Molecular weight LC-MS; MALDI-TOF MS
Peptide mapping RP-HPLC-UV
Sequence coverage LC-MS/MS
N-terminal/C-terminal sequence identification LC-MS/MS
Disulfide bond identification LC-MS/MS
N-glycosylation site identification LC-MS/MS
N-glycan analysis HILIC-HPLC/MS
PTM analysis LC-MS/MS
Higher order structure CD
pI icIEF
Charge variants icIEF;IEC-HPLC
Amino acid analysis RP-HPLC-UV/FLD
Purity & quantitation Purity SDS-PAGE; CE-SDS; SEC-HPLC
Quantitation UV; Bradford; Lowry; BCA; ELISA; AC-HPLC(protein A)
Impurity Host cell DNA(HCD) qPCR
Host cell Protein(HCP) ELISA
Residual Protein A ELISA
Safety & Stability Bacterial Endotoxins Gel-clot tech.; Chromogenic tech.
Bioburden Membrane filtration
Sterility Membrane filtration
Abnormal toxicity  Animal test (mouse)
Mycoplasma PCR
Subvisible particulate matter Light obscuration test
Visible particles Visual inspection
Tm DSC

Case 1: Accurate MW Determination by LC-MS

  • Case sample: Antibody
  • LC method: RP-HPLC column, 28 min Gradient
  • MS mode: TOF MS
  • Data analysis: PeakView

Figure 1. Original mass spectrum

Figure 2. Deconvoluted mass spectrum

Figure 3. Deconvoluted mass spectrum after deglycosylation

Case 2: Glycan analysis by UHPLC-MS

  • Sample: Antibody
  • Sample type: solution in PBS
  • LC method: HILIC-HPLC column, 65 min Gradient
  • Scan Type: TOF MS
  • Reagent Kits from Waters


Figure 4. N-glycan types of antibodies (Trends in biotechnology, Volume 34, Issue 10, p835–846)


Figure 5. Total ion chromatogram of N-glycan by LC-MS method

Name Formula MW(labled) Charge Theoretical MW Experimental MW PPM RT(min) Intensity
A1   1424.6 +2H 713.3 713.3 0.3 12.11 41858
M3   1221.5 +2H 611. 611. 0.9 12.11 1630
F(6)A1   1570.6 +2H 786.33 786.33 0.6 13.5 114406
G0
C67H105O37N9
1627.7 +2H 814.84 814.84 0.8 14.02 2714
G0F
C73H115O41N9
1773.7 +2H 887.87 887.87 0 15.26 2E+06
G2-2SA
C101H159O63N11
2535 +2H 1268.5 1268.5 -3.2 15.29 101
M5   1545.6 +2H 773.81 773.81 0 16.51 101
G1a/G1b
C73H115O42N9
1789.7 +2H 895.86 895.86 -0.8 16.82/17.23 446
F(6)A1G(4)1   1732.7 +2H 867.35 867.35 0.5 16.85 56477
G0F-GN
C81H128O46N10
19.8 +2H 989.41 989.41 -1 17.54 4359
G1Fa/G1Fb
C79H125O46N9
1935.8 +2H 968.89 968.89 -0.6 17.95/18.42 6666
G2
C79H125O47N9
1951.8 +2H 9.89 9.89 -0.1 19.93 5205
G1F-GN
C87H138O51N10
2138.9 +2H 1070.4 1070.4 -0.6 20.19 40
G1F-SA
C90H142O54N10
2226.9 +2H 4.4 4.4 -0.5 20.27 29
G2F
C85H135O51N9
2097.8 +2H 1049.9 1049.9 -0.7 20.99 1898
G2-SA
C90H142O55N10
2242.9 +2H 1122.4 1122.4 -1.9 21. 1037
F(6)M4A1G(4)1Ga(3)1
  2056.8 +2H 1029.4 1029.4 2.1 22.23 647
G2F-GN
C93H148O56N10
2301.9 +2H 1152 1152 -0.5 22.94 583
G2F-SA
C96H152O59N10
2389.9 +2H 1196 1196 0.1 23.03 78
G2F-2SA
C107H169O67N11
2681 +2H 1341.5 1341.5 -0.7 25.26 3239


Figure 6. 20+ N-glycan types identified in the table