Protein Analytical Service

Protein Analytical Service

With world-class analytical techniques, GenScript 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. GenScript ProBio's experts have expertise to take both recombinant proteins and monoclonal antibodies, from research and product development through characterization and quality control tests. Services are available for both standalone analytical assays and broad testing programs, to meet your specific needs.

Key Features

• Comprehensive breadth and innovative technologies
• Flexible services, available for both standalone analytical assays and broad testing programs

• Service Specifications
• Case Studies

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	cIEF; icIEF
	Charge variants	IEC-HPLC
	Amino acid analysis	RP-HPLC-UV/FLD
Purity & quantitation	Purity	SDS-PAGE; CE-SDS; iCE-SDS; SEC-HPLC
	Quantitation	UV; Bradford; Lowry; BCA; ELISA; AC-HPLC(protein A)
Impurity	Host cell DNA(HCD)	qPCR; SYBR green method
	Host cell Protein(HCP)	ELISA
	Residual Protein A	ELISA
Safety & Stability	Bacterial Endotoxins	Tal reagent
	Microbial limit	Membrane filtration
	Sterile examination	Membrane filtration
	Abnormal toxicity	Animal test (mouse)
	Mycoplasma	PCR
	Insoluble particles	Light obscuration test
	Visible particles	Lamp test
	Denaturation Temperature	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

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