Protein Analytical Service

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With world-class analytical techniques, ProBio is proud to provide a wide range of analytical testing services to ensure product quality and consistency during all stages of drug discovery and development. ProBio's experts have expertise to take both recombinant proteins and monoclonal antibodies, from research and product development through characterization and quality control tests.

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
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

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