KRIBIOLISA Pichia Pastoris (Wide Coverage) HCP ELISA

Name: KRIBIOLISA Pichia Pastoris (Wide Coverage) HCP ELISA
SKU: KBBP02
Availability: InStock

SKU: KBBP02

Rs 106,900.00

Immunoassay for the quantitative measurement of P.pastoris Host Cell Proteins as a contaminant in various biological preparations.
Other features include:
– Standardization and Reproducibility
– Lot to Lot consistency
– Accuracy and Precision to add confidence to your results
– Validated against seven points for a gold ring standard quality ELISA designed as per the US FDA guidelines for bioassays, a marker for Krishgen quality.
– Long shelf life and break-apart wells

Availability: 3 – 4 Weeks | Pack Size: 1 x 96 wells, break-apart

Introduction:
A variety of proteins and pDNA which are used as therapeutic agents in humans and animals are produced through recombinant expression in P.pastoris. The manufacturing and purification process of these products tends to leave the potential for contamination by Host Cell Proteins (HCPs) from P.pastoris which may result in adverse toxic or immunological reactions that ultimately affect the efficacy of the therapeutic agent. The simple, objective and semi quantitative ELISA is a highly sensitive method that aids in purification process development, process control, quality control and product release testing optimally.

Intended Use:
This KRIBIOLISA Pichia Pastoris HCP ELISA is a generic kit intended in determining the presence of P. pastoris Host Cell Proteins contamination in various biotechnology products that are manufactured through recombinant expression in P. pastoris. The kit has been validated successfully for testing of in process and final product HCPs in variety of products regardless of growth and purification process.

Principle:
This assay is based on the Sandwich ELISA procedure. Samples containing P.pastoris HCPs are reacted with already coated affinity purified capture anti-P.pastoris antibody and bind to them. Plates are washed with wash buffer to remove unbound reactants. Anti P.pastoris-HRP conjugate is added which results in the formation of a sandwich complex of solid phase antibody-HCP-enzyme labeled antibody. The wells are washed to remove any unbound reactants as per the wash procedure. The substrate Tetra Methyl Benzidine is then reacted. The amount of hydrolyzed substrate is read on a microtiter plate reader and it is directly proportional to the concentration of P. pastoris HCPs present.

If you have published a paper by using any of our ELISA since 01/01/2023, kindly fill out the “Krishgen Publication Reward Application Form” with complete information and send it by at email: kbiinfo@krishgen.com, with the subject “Krishgen Publication Reward”. We will get back to you with the Amazon / Krishgen Credit Reward after we confirm it ASAP!

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Additional information

Pack Type	ELISA 96 wells
Species Reactivity	PPastoris cells
Sample Type	Cell Culture Supernatant and other biological samples
Assay Range	2-200 ng/ml
Sensitivity	0.5 ng/ml
Interference	Preparations of the known factors were assayed for interference. No significant interference was observed.
Specificity	The assay recognises host cell proteins including low and high molecular weight proteins.
Principle Of Assay	Double Antibody, Sandwich ELISA
Detection Method	Absorbance measured at 450nm, Colorimetric
Conjugate-Enzyme Reaction	Uses HRP enzyme conjugate with TMB chromogenic substrate for color development
Quality Certification	Validated as per KRISHGEN's Gold Ring 7-Point Program
KRISHGEN Mandate	High Performance Assays at Affordable Prices
Regulatory Status	Research Use Only.
Shipping Temperature	Room Temperature / 2-8 degrees Celsius
Storage Temperature	if all the wells in the kit are not being used, store the unused wells in the foil pouch containing the desiccant pack, The kit and its components are to be stored as indicated in the IFU (instructions for use). To ensure quality of your results, well sealed and store at the indicated temperature.
ELISA Type	Double Antibody, Sandich ELISA
ELISA Interpretation	on request, Quantitative Results Intepretation (% Binding/OD450), Software (MS Excel) available, to support interpretation of results.
Shelf Life Available	8 -10 months at time of shipping
Alternate Names / Synonyms	Pichia Pastoris HCP; ppastoris; host cell proteins
Research Area	Biosimilars; Monoclonal Antibodies; small molecules; therapeutic drug monitoring assays
Disclaimer	The data indicated herein with specifications are changed from time to time at time of production. Please refer the most current IFU (instructions for use) accompanying the kit.

Protocol

Assay Procedure:
Bring all reagents to room temperature prior to use. It is strongly recommended that all standards and samples be run in duplicate or triplicate. A standard curve is required for each assay.
1. Pipette 100 ul of Standards or Samples into the respective wells.
2. Cover the plate and incubate at 37 Degree Celcius for 2 hours 30 minutes.
3. Aspirate and wash plate 4 times with Wash Buffer (1X) and blot residual buffer by firmly tapping plate upside down on absorbent paper. Wipe of any liquid from the bottom outside of the microtiter wells as any residue can interfere in the reading step.
4. Add 100 ul of Anti-P.pastoris:HRP Conjugate into each well.
5. Cover the plate and incubate at 37 Degree Celcius for 1 hour.
6. Aspirate and wash plate 4 times with Wash Buffer (1X) and blot residual buffer by firmly tapping plate upside down on absorbent paper. Wipe of any liquid from the bottom outside of the microtiter wells as any residue can interfere in the reading step.
7. Add 100 ul of TMB Substrate in each well.
8. Incubate the plate at 37 Degree Celcius for 15 minutes in dark. DO NOT SHAKE or else it may result in higher backgrounds and worse precision. Positive wells should turn bluish in color.
9. Pipette out 100 ul of Stop Solution. Wells should turn from blue to yellow in color.
10. Read the absorbance at 450 nm with a microplate reader within 30 minutes.