Manufacturing Process and Principle of Radioimmunoassay Kit ShinJin Medics S.A. Diagnostic Development Lab

Content Ⅰ. Design Process Ⅱ. Manufacturing Process Ⅲ. Quality Control Process

I. Design Process

Basic concept of Radioimmunoassay Tracer Capture

Development of Raw materials                                                               Monoclonal Antibody development Polyclonal Antibody development

                                                                                                                                                                                                                                                                            

Development of Radioimmunoassay                                                          Matching Test Performance Test Stability Test Evaluation Step Tracer iodination Capure coating Matching Test Basic clinical Test Sensitivity Test Specificity Test Precision Test Recovery Test Linearity Test Hook effect Test Tracer stability Test Tube stability Test Standard stability Test External Evaluation Internal

II. Manufacturing Process

Immunotube Manufacturing Methods 1. Coating 2. Blocking 3. Fixing

Coating Instrument

Blocking / Fixing Instrument

Iodination Methods Iodination of antibodies and antigens is a straighforward and effective method of labeling I125 is normally used for most immunochemical analyses The decay of I125 yields low-energy gamma and X-ray radiation , easy to detect. The 60-day half-life of I125 is convenient compromise between effective shelf life and waste disposal.

Iodination Methods

Bolton-Hunter reagent 3-(4-Hydroxyphenyl)propionic acid N-hydroxysuccinimide ester  React with I125 React with protein, petide

Target material of Iodination Protein Small molecule 1. Steroid 2. Thyroid

Process of Protein Iodination 1. Iodine 125 Tagging I125 + = I125

Purification of Protein-I125

Steroids Iodination Structure of Steroids Kinds of Steroids 1. Estradiol (E2) 2. Estriol (E3) 3. Progesterone 4. Testosterone 5. Cortisol

Process of Steroids Iodination Materials 1. Steroid 2. Histamine 3. I125

Method 1. Steroid activation 2. Histamine activation 3. Conjugation of Steroid and Histamine-I125 4. Purification of Steroid and Histamine-I125 by TLC (Thin layer Chromatography) method

III. Quality Control Process

QC Process Raw Materials QC Tube QC Tracer QC Semi-Assembled QC Coating Raw Materials QC Tracer Raw Standard Raw Materials QC Coating Buffer Blocking Buffer Fixing Buffer QC QC of coated Tube (sensitivity, specifity) QC of Iodinated Tracer (sensitivity, specifity) Before Assembled, All components tested (Tube and Tracer and Standard etc) After Assembled, kits tested

LLHL Effect (Low Lower, High Lower) Tube QC Background LLHL Effect (Low Lower, High Lower) Con. Cpm Con. Cpm After Damage

LHHL Effect (Low Higher, High Lower) Tracer QC Background LHHL Effect (Low Higher, High Lower) Con. Cpm Con. Cpm After Damage Hinge point

Example of damaged Tracer Hinge point

Conclusion Capture and tracer selected and developed through monoclonal Ab and polyclonal Ab The immunotube is manufactured through coating, blocking, fixing phase. Tracer is manufactured through I125 coupling by oxidation method.

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