YY/T 0878.3-2019Test for complement activation of medical devices - Part 3: Assay for the product of complement activation (C3a and SC5b-9) (English PDF)

Foreword

YY/T 0878 "Medical Device Complement Activation Test" includes the following parts.

--- Part 1. Serum full complement activation;

--- Part 2. Serum bypass pathway complement activation;

--- Part 3. Determination of complement activation products (C3a and SC5b-9). This part is the third part of YY/T 0878.

This part is drafted in accordance with the rules given in GB/T 1.1-2009.

Introduction

The selection strategy for the blood compatibility test of medical devices/materials is given in GB/T 16886.4. This part of YY/T 0878 is in vitro.

The specific test method for complement activation products can be used as a supplement to the medical device/material complement activation test in GB/T 16886.4.

After the medical device/material is in contact with blood, it may activate the complement system in the blood, and the production of effector molecules may cause the body to be severely acute and chronic.

The reaction also affects the blood compatibility of the medical device/material. C3a is a small molecule formed by the activation of complement C3 molecule and can be mediated.

Acute inflammatory response of the body. SC5b-9 is a co-terminal product C5b-9 produced by activation of the complement system and formation of S protein in serum A hydrophilic, non-lytic cell-active macromolecular fragment or a macromolecular fragment without lytic cell membrane activity. By detecting C3a molecules or The content of SC5b-9 molecule can reflect the activation of the complement system in serum. In vitro supplement as described in this section of YY/T 0878 A method for determining a body activation product that can be used to assess whether a medical device/material has a potential complement activation.

1 Scope

This section of YY/T 0878 specifies methods for the determination of complement activation products (C3a and SC5b-9) in the blood by enzyme-linked immunosorbent assay.

This section applies to the evaluation of the activation potential of medical devices/materials in vitro.

The test system established in this section is applicable to human serum or anticoagulated plasma. If animal blood is used, the test is carried out after demonstrating its suitability.

2 Normative references

The following documents are indispensable for the application of this document. For dated references, only dated versions apply to this document. For undated references, the latest edition (including all amendments) applies to this document.

GB/T 16886.1 Biological evaluation of medical devices - Part 1. Evaluation and testing in the process of risk management

GB/T 16886.4 Biological evaluation of medical devices - Part 4. Selection of test for interaction with blood

GB/T 16886.12 Biological evaluation of medical devices - Part 12. Sample preparation and reference materials

3 Terms and definitions

The terms and definitions defined in GB/T 16886.1, GB/T 16886.4 and GB/T 16886.12 apply to this document.

4 Test principle (taking C3a as an example)

C3a levels in serum or anticoagulated plasma were determined by double antibody sandwich (ELISA). Coating micropores with purified rat C3a antibody The plate is made into a solid phase antibody, and then human serum or anticoagulated plasma is sequentially added to the microwell of the coated monoclonal antibody. Horseradish peroxidase (HRP)-labeled C3a antibody, forming an antibody-antigen-enzyme-labeled antibody complex, after thorough washing, adding substrate tetramethylbenzidine (TMB) color development. TMB is converted to blue under the catalysis of HRP and converted to the final yellow color by the action of an acid. The depth of the color There was a positive correlation between the concentration of C3a in serum or anticoagulated plasma. The absorbance (OD value) was measured at 450 nm using a microplate reader and passed the standard.

The curve calculates the concentration of human C3a in serum or anticoagulated plasma.

5 Reagents and materials