The production of the main components of luminescent immunoassay reagents includes steps such as coating reaction plates, preparation of markers, configuration of various solutions, freeze-drying, and subpackaging; and two quality control processes of semi-finished product inspection and finished product inspection to ensure its The quality is as specified.
1. Preparation of solid-phase carrier (because the coated carriers used by different products are very different, the standard 96-well microwell reaction plate is used as an example to describe here)
1) Preparation of coated plates
Prepare coated boards that have passed the inspection, and record the batch number, quantity, and status identification.
Quality control items: size, appearance, packaging.
(2) Preparation of coating solution
Prepare the coating buffer, add the coated antibody or antigen to the working concentration, and mix well to form the required coating solution. The working concentration of the coating solution should be used within the specified time.
Quality control items: coating buffer formula, pH value, coating composition.
(3) Coating of the coated board
The coating solution is added to the coated plate according to the process requirements. Record the number of coated plates coated.
Quality control items: coating volume, temperature, time, process monitoring.
(4) Preparation of plate washing working solution (according to the process requirements of each unit, plate washing is not required)
Prepare the plate washing working solution according to the formula.
Quality control items: plate washing working solution formula, pH value.
(5) Preparation of blocking solution
Prepare blocking solution according to recipe.
Quality control items: blocking solution formula, pH value.
6) Washing and sealing
After the coating is completed, remove the coating solution in the well, wash the plate with the plate washing working solution (according to the process requirements of each unit, the plate can not be washed), and then add the blocking solution.
Quality control items: closed volume, temperature, time, process monitoring.
Quality inspection items: check the uniformity of the package before sealing.
(7) Drain
After sealing the reaction plate, drain the liquid in the well.
Quality control items: process monitoring.
(8) dry
The reaction plate should be dried according to the requirements of the process.
Quality control items: temperature, humidity, time, process monitoring, etc.
(9) Sealed packaging
Seal the dried reaction plate with an aluminum foil bag, and put a desiccant inside (according to the process requirements of each unit, it can not be put).
Quality control items: sealing performance, labeling and expiration date, etc.
(10) Reaction plate (semi-finished product) inspection
Sampling inspection is carried out on the bagged and sealed reaction plates for appearance, intra-plate variation, and inter-plate variation.
2. Dropping process
(1) Preparation of enzyme conjugates (according to the actual situation of each product, this step may not be carried out)
Use the conventional sodium periodate-ethylene glycol method to label the relevant antibody (or antigen) with horseradish peroxidase (or other enzymes), and the enzyme-labeled antibody (or antigen) should be added with an appropriate protective agent and stored in low temperature.
Quality control items: marking method, process control.
(2) Identification of enzyme conjugates
①Functional experiment
After diluting the enzyme conjugate with enzyme diluent, it is used for the dropwise dispensing of the product, and the result should meet the quality standards of the relevant kit.
② Stability
Dilute the enzyme conjugate with enzyme diluent, and conduct a thermal stability test at 28°C. The dropping results should meet the quality standards of the relevant kits.
(3) Enzyme Conjugate Diluent
Prepare according to the formula of enzyme conjugate diluent, store at 28°C, and use within the specified time.
Quality control items: enzyme conjugate dilution formula, pH value.
(4) Dosing of the working concentration of the enzyme conjugate
Take the enzyme conjugate, dilute it to different concentrations with the enzyme conjugate diluent, and use the prepared reaction plate for dropwise dosing. Measure a series of standard products and corresponding quality control products to determine the optimal working concentration of the enzyme conjugate for the system.
(5) Preparation of enzyme conjugate working solution
Mix the required amount of enzyme conjugate and enzyme conjugate dilution according to the drop concentration.
Quality inspection items: inspection before subpackaging, using the matching reaction plate for inspection, appearance, sensitivity, measured value of quality control products, and quantitative products should be used for linear inspection of calibrator.
(6) Subpackage of enzyme conjugate working solution
Pack the enzyme conjugate working solution according to the process requirements.
Quality control items: Confirm the reagent name, batch number, quantity, subpackage volume, and sealability after packaging before subpackaging.
(7) Inspection of enzyme conjugate working solution (semi-finished product)
Sampling inspection of the enzyme conjugate working solution after packaging, appearance, filling volume, sensitivity, linearity of the dose-response curve of the calibrator, and the measured value of the quality control product.
3. Preparation of calibrators, negative/positive controls or quality controls
(1) diluent
Prepare according to the formula of the diluent, store at 28°C or -20°C, and use within the validity period.
Quality control items: diluent formula, pH value.
(2) Preparation of calibrator, negative/positive control or quality control
The preparation of calibrators, negative/positive controls or quality control products should be traceable in quantity and value, and can be prepared with reference to national standard products, World Health Organization standard products or other grades of standard materials.
Quality inspection items: pre-packaging inspection, accuracy, dose-response curve linearity (quantitative products), measured value of quality control products.
(3) Aliquoting of calibrators, negative/positive controls or quality controls
Dispense calibrators, negative/positive controls or quality controls according to process requirements.
Quality control items: Confirm the reagent name, batch number, quantity, and aliquot volume before subpackaging.
(4) Inspection of calibrator, negative/positive control or quality control (semi-finished product)
Carry out sampling inspection on the calibrators, negative/positive controls or quality control products after packaging, appearance, filling volume, accuracy, dose-response curve linearity (quantitative products), and measured values of quality control products.
4. Preparation of Chemiluminescent Substrates
(1) Substrate buffer
Prepare according to the formula of substrate buffer, store at 28°C, and use within the validity period.
Quality control items: substrate buffer formula, pH value.
(2) Preparation of chemiluminescent substrates (oxidants and luminescent agents)
Add the corresponding oxidizing agent and luminescent agent to the substrate buffer according to the formula of oxidizing agent and luminescent agent respectively
Quality control items: oxidizing agent and luminescent agent formula.
Quality inspection items: inspection before packaging, background, luminous intensity.
(3) Subpackaging of chemiluminescent substrates (oxidants and luminescent agents)
Pack chemiluminescent substrates (oxidants and luminescent agents) according to process requirements.
Quality control items: Confirm the reagent name, batch number, quantity, subpackage volume, and sealability after packaging before subpackaging.
(4) Inspection of chemiluminescence substrate (semi-finished product)
The chemiluminescent substrates after packaging are sampled and inspected for appearance, filling volume, background, sensitivity, and luminous intensity.
5. Preparation of Europium Labels
For different labeled biological raw materials, determine different labeling preparation processes, including the ratio of europium-DTTA to labeled biological raw materials, labeling temperature and labeling time, calculation standards for labeling yield, etc. In the actual operation process, it is required to strictly follow the standard operating procedures
6. Freeze drying
All kinds of freeze-dried products need to establish their own freeze-drying process. The appearance of the freeze-dried products should be a loose powdery solid with a certain shape, and it can be completely and quickly reconstituted into a clear and transparent liquid.
7. Packing, light inspection and labeling
The dispensing volume is measured by weight loss weighing method, and the dispensing volume is controlled after converting the mass into volume. Light inspection is to visually inspect the color of each component, the filling volume, and whether there are turbidity and impurities.
8. Packaging
According to the requirements of the kit packaging standard operating procedures and instructions, it is packaged in the form of assembly line operation. When packaging, the product name, batch number, and loading quantity should be strictly checked, and the quantity of each material should be carefully checked, and re-checked before closing the box.
