HTRF CHO Host Cell Protein Detection Kit, 10,000 Assay Points

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HTRF CHO Host Cell Protein Detection Kit, 10,000 Assay Points

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The CHO Host Cell Protein kit is designed for the quantitative assessment of Chinese Hamster Ovary (CHO) cell contaminants in bioprocess operations.

For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption and disposal requirements under European REACH regulations (EC 1907/2006).

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Feature	Specification
Application	Protein Quantification
Sample Volume	16 µL

The CHO Host Cell Protein kit is designed for the quantitative assessment of Chinese Hamster Ovary (CHO) cell contaminants in bioprocess operations.

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Product Variants

Unit Size: 500 Assay Points

Part #:

64CHOPEG

Unit Size: 10,000 Assay Points

Part #:

64CHOPEH

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Product information

Overview

The CHO Host Cell Protein (HCP) kit measures contaminants originating from the CHO cells used to manufacture therapeutic antibodies. The kit can be used for the quantification of CHO HCP proteins in routine bioprocess operations, from the completely raw harvest material to the final product. The simple and robust procedure benefits from increased throughput compared to ELISA.

Specifications

Other Specifications

Application	Protein Quantification
Automation Compatible	Yes
Brand	HTRF
Detection Modality	HTRF
Product Group	Kit
Sample Volume	16 µL
Shipping Conditions	Shipped in Dry Ice
Target Class	Biomarkers
Technology	TR-FRET
Therapeutic Area	Cardiovascular Infectious Diseases Metabolism/Diabetes NASH/Fibrosis Neuroscience Rare Diseases
Unit Size	10,000 Assay Points

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How it works

Assay principle

The CHO Host Cell Protein (HCP) assay is a sandwich immunoassay. The assay uses a specific and qualified anti-CHO HCP polyclonal antibody pool, labelled with Eu3+-Cryptate and t d2. The specific signal is proportional to the concentration of CHO HCP present in the sample. This homogeneous assay requires only a single reagent addition (by premixing the detection reagents), and an overnight incubation, for the quantitative detection of CHO HCP.

Assay protocol

The CHO HCP assay protocol, using a 384-well small volume white plate, is described on the right. 16 µL of sample or standard are dispensed directly into the detection plate for detection by HTRF® reagents. The antibodies labeled with the HTRF donor and acceptor can be pre-mixed and added in a single dispensing step, to further streamline the assay procedure. This protocol requires only 1 overnight incubation period.

Assay details

HTRF CHO HCP assay specifications

Sample volume	16 µL
Final assay volume	20 µL
Time to results	Overnight at RT
Detection limit	0.8 ng/mL
Dynamic range	3.6 to 400 ng/mL
Sample compatibility	From raw harvest material to the final biotherapeutic product

HTRF CHO HCP assay specifications

Analytical performance

Precision

Intra-assay (n=24)

Sample	Mean [CHO HCP] (ng/mL)	% CV
1	10	6%
2	29	3%
3	97	3%
4	306	4%
Each of the 4 samples was measured 24 times, and the %CV on concentration was calculated for each sample.

Inter-assay (n=4)

Sample	Mean [CHO HCP] (ng/mL)	%CV
1	10	9%
2	29	5%
3	97	4%
4	292	5%
Each of the 4 samples was measured in 4 independent experiments by 4 different operators, and the concentration % CV was calculated for each sample.

Antigen spike and recovery

Sample	[CHO HCP Standard] (ng/mL)	[CHO-DG44 HCP Sample] added (ng/mL)	Expected [CHO HCP] (ng/mL)	Measured [CHO HCP] (ng/mL)	% Antigen recovery
1	13.51	7.04	20.55	21.38	104%
2	13.51	14.07	27.58	27.92	101%
3	13.51	23.38	36.89	41.25	112%
4	45.55	26.04	71.59	69.45	97%
5	45.55	45.06	90.61	94.08	104%
6	45.55	88.02	133.57	137.62	103%
7	95.11	45.06	140.17	150.64	107%
8	95.11	88.02	183.13	202.40	111%
9	95.11	161.56	256.67	279.14	109%

Three different levels of CHO HCP Standard were independently mixed with three different sample concentrations (cell culture supernatant from a biomanufacturing process using the CHO-DG44 cell line). The total measured concentrations were compared to the theoretical ones, and expressed as % Antigen recovery (acceptance criteria: 85-115%).

CHO-S HCP sample
Dilution Factor	Expected [CHO HCP] (ng/mL)	Measured [CHO HCP] (ng/mL)	% Recovery
Neat	426	426	100%
2	213	226	106%
4	106	111	104%
8	53	57	106%
16	27	28	107%
32	13	13	98%
64	7	7	101%
128	3	4	106%
Samples were from cell culture supernatant from a biomanufacturing process using the CHO-S cell line diluted in the kit's Diluent 5. The recovery % obtained from these experiments show the good dilutional linearity of the assay (dilution test acceptance criteria: 80-120%)

CHO-K1 HCP sample
Dilution Factor	Expected [CHO HCP] (ng/mL)	Measured [CHO HCP] (ng/mL)	% Recovery
Neat	344	344	100%
2	172	173	100%
4	86	88	102%
8	43	45	105%
16	22	24	111%
32	11	13	119%
Samples were from cell culture supernatant from a biomanufacturing process using the CHO-K1 cell line and diluted in the kit's Diluent #5. The recovery % obtained from these experiments show the good dilutional linearity of the assay (dilution test acceptance criteria: 80-120%)