Protein/Ab characterization

Key Features

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
Purity & quantitation	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

Figure 1. Original mass spectrum

Figure 2. Deconvoluted mass spectrum

Figure 3. Deconvoluted mass spectrum after deglycosylation

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