0 item(s)
View cart
You have no items in your shopping cart.
Product Description
A 96-well solid-phase, double-antibody sandwich one-step ELISA for quantitative detection of residual host cell proteins (HCPs) from Pichia pastoris (P. pastoris) strains (e.g., GS115, X33). The kit uses an affinity-purified sheep polyclonal anti-P.pastoris HCP capture antibody coated on microtiter strips and an HRP-conjugated sheep anti-P.pastoris HCP detection antibody added simultaneously with calibration standard or sample. TMB/H2O2 is used as the substrate (blue product at ~655 nm; stopped to yellow product with maximum absorbance at 450 nm). Sample concentrations are calculated from a 4-parameter logistic (4-PL) dose–response calibration curve using OD450 (with 620–650 nm reference subtraction). The kit is intended for research use only.
Technical Specifications
| Parameter | Details |
|---|---|
| Assay format | One-step double-antibody sandwich ELISA (pre-coated microtiter strips) |
| Plate format / Kit size | 96 tests per kit (8-well × 12 strips) |
| Linear range | 2–200 ng/mL (4-PL fit, R² ≥ 0.990) |
| LLOQ | 2 ng/mL |
| Typical calibration curve | Standards at 0, 2, 10, 20, 50, 100, 200 ng/mL; example OD450-620nm average: 0 ng/mL ~0.078; 2 ng/mL ~0.114; 10 ng/mL ~0.270; 20 ng/mL ~0.434; 50 ng/mL ~0.761; 100 ng/mL ~1.233; 200 ng/mL ~1.704 |
| Specificity / Cross-reactivity | No cross-reactivity with E. coli, CHO and Sf9 cells; <1% cross-reactivity with Saccharomyces cerevisiae and Hansenula polymorpha |
| Detection wavelength | Read at 450 nm with reference 620–650 nm (OD450 nm - reference) |
| Assay incubation times | Sample/HRP incubation 3 hours at room temperature (600 rpm recommended); TMB substrate incubation 10 minutes; post-stop incubation/read within recommended time |
| Recommended sample pH | pH 6.5–8.5 |
| Recommended sample dilution | Typically ≥10-fold dilution recommended; method validation recommended to determine optimal dilutions |
| Read time window | Complete reading within 10 minutes after stopping (acceptable up to 30 minutes) |
Features
- One-step simultaneous detection: Calibration standard (or sample) and HRP-conjugated detection antibody are added simultaneously to reduce handling steps.
- High sensitivity: Low limit of quantitation (LLOQ) of 2 ng/mL across a 2–200 ng/mL linear range with 4-PL fitting (R² ≥ 0.990).
- High specificity: No cross-reactivity with common non-P.pastoris hosts (E. coli, CHO, Sf9); minimal cross-reactivity (<1%) with closely related yeasts.
- Ready-to-use pre-coated strips: Microtiter strips are pre-coated with affinity-purified sheep anti-P.pastoris HCP antibody and packaged with desiccant for convenience.
- Complete kit: Includes calibration standard, diluent, wash buffer concentrate, HRP conjugate, TMB substrate, stop solution and sealing film for full assay workflow.
Applications
- Quantitation of residual host cell proteins (HCPs) in products manufactured with P. pastoris (e.g., GS115, X33)
- Testing in-process samples, harvested bulk, drug substance and drug product
- HCP monitoring during downstream purification and process development
- Research applications for bioprocess development and analytical method validation
Kit Contents
P.pastoris HCP Calibration Standard (lyophilized)2 bottles; reconstitute with 500 µL Reconstitution Solution. Reconstituted: 2-8 °C up to 7 days or < -20 °C up to 60 days
Anti-P.pastoris HCP Microtiter Strips (pre-coated)8 wells × 12 strips; store in vacuumed bag with desiccant. Opened: 2-8 °C up to 60 days
Reconstitution Solution1 × 1.5 mL; room temperature equilibration for use; used to dissolve calibration standard
Diluent2 × 25 mL; room temperature; used for dilution of standards, HRP(100×) and samples
Wash Buffer Concentrate (10×)2 × 25 mL; dilute 1:10 with ultra-pure water before use
Anti-P.pastoris :HRP (100×)1 × 120 µL HRP-conjugated affinity purified sheep antibody; centrifuge before use; dilute 100× in Diluent immediately prior to use
TMB Substrate1 × 12 mL; sealed, keep away from light; equilibrate to room temperature (~20 min) before use
Stop Solution (1 M HCl)1 × 6 mL; corrosive—avoid contact with eyes, skin and clothing
Sealing Film3 pieces; use to cover strips during incubation
* Total: 96 tests/Kit | Method: One-step ELISA (double-antibody sandwich)
Attention
• Research Use Only (not for clinical diagnostic use).
• Store kit at 2–8 °C; check expiration date on labels.
• Equilibrate reagents to room temperature (~20 min) before use.
• Prepare fresh 1× Anti-P.pastoris:HRP by diluting 100× stock in Diluent immediately before use; centrifuge HRP(100×) before dilution.
• Reconstituted calibration standard solutions and anti-P.pastoris HCP antibody solution are recommended for single use due to instability.
• Avoid samples containing sodium azide (NaN3) because it inactivates HRP.
• Sample pH must be between 6.5 and 8.5; adjust or buffer samples accordingly.
• Inactivate proteolytic enzymes in samples to prevent degradation of HCPs.
• Do not mix reagents from different lot numbers.
• Avoid drying wells before substrate addition; perform recommended wash steps (5×) and remove residual liquid.
• TMB should be colorless before use; discard if colored.
• Stop Solution is 1 M HCl — use appropriate PPE and avoid contact with skin/eyes/clothing.
• Read plate within 10 minutes after stopping for most stable results (acceptable up to 30 minutes).
• Use sterile, ultra-pure water; if Wash Buffer or Diluent is cloudy, heat to 37 °C until clear.
Quality Management & Certifications
Quality System
Download QMSQMS (ISO, GMP)
Download CertificateQuality Advantages
View ReportQuality Control Process
Download ProcessTechnical Resources & Downloads
Frequently Asked Questions (FAQ)
Q1: How do I reconstitute the calibration standard?
Add 500 µL Reconstitution Solution to a lyophilized calibration standard vial, invert gently and let stand ~5 minutes. Combine vials if multiple are used and mix gently. Prepare serial dilutions as described (ST1 = 200 ng/mL, ST2 = 100 ng/mL, etc.).
Q2: What is the assay sensitivity (LLOQ) and linear range?
LLOQ is 2 ng/mL. The validated linear range is 2–200 ng/mL using a 4-parameter logistic (4-PL) fit with R² ≥ 0.990.
Q3: What sample dilution should I use?
Typically start with at least a 10-fold dilution for in-process, harvest or drug substance samples. Conduct method validation and spike-recovery experiments to determine optimal dilutions for your matrix.
Q4: How should I store reconstituted standards and opened components?
Reconstituted standard: 2–8 °C for up to 7 days for short-term use; for long-term storage keep below -20 °C for up to 60 days. Opened microtiter strips: store in bag with desiccant at 2–8 °C up to 60 days.
Research Use Only
✔
Research Use Only (RUO); Not for clinical diagnostic use – This product is intended for laboratory research purposes only and not for clinical or regulatory diagnostic use.
✔
Not intended for use in USDA or FDA regulated diagnostic testing or official compliance testing.
When can I expect my order to ship?
Most orders are filled and shipped within 2-3 business days from the time they are received.
Our standard shipping usually take 2-5 days.
We also provide express shippping for time-sensitive deliveries.
Email contact@biofargo.com if you have any requirements.
