CHO HCP ELISA Kit,Absin_specification/price/image

Product Detail
Company Profile

Product Specification

Usage

1. Reagent preparation:
1. PBST 0.05% 15ml 20 × PBST 0.05%, dilute in ddH2O, dilute to 300ml.
2. 1.0% BSA Dissolve 1g of BSA in the bottle in 100ml PBST 0.05%, mix thoroughly until completely dissolved, and store at 2-8 °C. The prepared dilution buffer is valid for 7 days. It is recommended to prepare it as needed.
3. Test solution 2μg/mL Take 48 μL of 0.5 mg/mL Anti-CHO HCP-HRP and dilute it in 11952 μL of 1% BSA to obtain a final concentration of 2μg/mL test solution.
4. Preparation of QC and CHOK1 HCP standards

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Tube number	Drug substance	Concentration (ng/mL)	Volume (μL)	Volume of 1% BSA (μL)	Total Volume	Final Concentration (ng/mL)
A	Standard	0.2 mg/mL	10	490	500	10000
B	A	10000	50	450	500	1000
S1	B	1000	50	450	500	100
S2	S1	100	300	100	400	75
S3	S2	75	200	175	375	40
S4	S3	40	150	350	500	12
S5	S4	12	200	200	400	6
S6	S5	6	200	200	3
NC	NA	NA	NA	200	200	0
QC	S1	100	50	200	250	20

2. Required instruments:

Consumables/Equipment	manufacturer	model
Microplate reader	Molecular Devices	Spectra Max M5, M5e, Or equivalent
Thermostatic mixer	Eppendorf	Eppendorf/5355, or equivalent
Vortex mixer	IKA	MS3 Digital, or equivalent

3. Experimental process
1. Prepare the reagents according to the above "reagent preparation".
2. Take 50μL of standard, sample and QC on the well plate, then add 100μL of test solution (2μg/mL), seal the labeled plate with a sealing membrane, place the labeled plate on the mixer, incubate at 500 rpm, 25 ± 3 ℃ for 2 hours.
3. Turn the microplate upside down in the water tank and discard the coating liquid. Take 300 μL of PBST 0.05% and wash the plate in microwells and discard the solution, and repeat the wash three times. Snap the board upside down on a clean paper towel and pat dry.
4. Add 100μL of TMB substrate to each well, seal the labeled plate, and incubate at 25 ± 3 ℃ in the dark for 15 minutes.
5. Add 100 μL of stop solution to each well.
6. Read the absorbance at the wavelength of 450/650 nm with the microplate reader.
7. Analyze the data by SoftMax or similar software, and draw the standard curve with four-parameter fitting regression model.

Note: If the concentration of HCP in the sample exceeds the upper limit of the standard curve, it needs to be properly diluted with dilution buffer prior to testing.

Theory

This method uses one-step enzyme-linked immunosorbent ELISA. Samples containing CHOK1 HCP can be simultaneously reacted with HRP-labeled goat anti-CHOK1 antibodies and anti-CHOK1 antibodies coated on enzyme plates to finally form a sandwich complex of solid phase antibodies-HCP-labeled antibodies. Unbound antigen antibodies can be removed by washing the plate. After the TMB substrate is added to the well to complete the reaction, the color development is stopped after the stop solution is added, and the OD or absorbance value of the reaction solution at 450/650 nm is read with a microplate reader. The OD value or absorbance value is proportional to the HCP content in the solution. From this, the HCP concentration in the solution can be calculated from the standard curve.

Description

Host Cell Protein (HCP) secreted by CHO (Chinese Hamster Ovary) host cells is likely to cause contamination of biological products. Failure to adequately remove contaminants in the early stage of drug development may lead to a decrease in drug efficacy or induce an immune response. The kit can quantitatively detect the residue of host cell protein in bioengineering pharmaceuticals with high sensitivity, optimize the process flow and ensure the safety of biological products.
Product parameters:
1. Sensitivity: 1ng/mL
2. Detection range: 3-100ng/mL
3. Precision: intra-batch coefficient of variation ≤ 10%, inter-batch coefficient of variation ≤ 15%
4. HCP coverage: more than 80%
5. Specificity: This kit is universal because it can specifically react with CHOK1 HCP without relying on the purification process

Composition

Serial number	form	Concentration	Specifications	Storage conditions
1	CHOK1 HCP Standard	0.5 mg/mL	70uL/tube	≤-20℃
2	Anti-CHO HCP-HRP	0.5 mg/mL	60uL/tube	≤-20 ℃, protected from light
3	TMB	NA	12mL/vial	2-8 ℃, protected from light
4	20 × PBST 0.05%	NA	15mL/vial	2-8℃
5	Stop liquid	NA	12mL/vial	Room temperature
6	Enzyme labeled plate sealing membrane	NA	3 sheets	Room temperature
7	BSA	NA	1g	2-8℃
8	High adsorption 96-well prediction enzyme plate	NA	1 piece	2-8℃

General Notes

The termination solution is 2M sulfuric acid, please handle it carefully to prevent splashes!

Instructions

This kit is used to quantitatively detect the residual content of CHOK1 host cell protein in a sample.

Storage Temp.

Components 1 and 2 are stored at ≤-20 °C; Components 5 and 6 are stored at room temperature; Components 3, 4, 7, 8 are stored at 2-8 °C. Valid for 12 months.

AntBio is a biotechnology group company dedicated to serving life sciences, aiming to help scientists accelerate research and improve work efficiency. AntBio provides comprehensive and high-quality reagent tools for basic research, drug development, and diagnosis, including research grade antibodies, proteins, biochemical reagents, and assay kits. These research tools are widely used in different segments of life science research. The group company currently consists of three brands, Absin, Starter-Bio and UA-Bio.