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CHO HCP ELISA Kit

CHO HCP ELISA Kit

Catalog Number: abs590005 Application: ELISA Brand: Absin
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$952 USD
$952 USD
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Product Details

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 2003
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.

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