Lateral Flow Assay Development

SERVICE NO: BOTYW0001

Lateral flow assay development (LFA) is a process of constructing a rapid detection system by utilizing the principle of specific antigen-antibody binding, with immunochromatography as the core. This process requires first clarifying the detection target, selecting and preparing highly specific antibodies/antigens, matching with markers such as colloidal gold and modifying them to enhance binding efficiency, followed by strip assembly and performance evaluation. Key considerations during the development include: ensuring the quality of reagents and materials (specificity/affinity of antibodies/antigens, stability of markers, and performance of chromatographic materials), precisely optimizing detection parameters (strictly controlling buffer pH, ionic strength, detection time, and temperature), comprehensively evaluating performance (including sensitivity, specificity, and stability tests), and standardizing production processes to ensure product quality consistency. This technology is widely applied, enabling the detection of pathogens (such as SARS-CoV-2), tumor markers, hormones, etc., in the biomedical field, and the rapid detection of harmful substances such as pesticide/veterinary drug residues, heavy metals, and microorganisms in food safety, providing effective solutions for rapid detection in multiple fields.

Lateral Flow Assay Development Condition

1.High specific antibody / antigen (prepared by hybridoma or genetic engineering technology, the affinity constant needs to reach more than 10 ? 9m), stable markers (such as colloidal gold with uniform particle size, microspheres with high fluorescence quantum yield), and suitable reaction buffer (pH value and ion strength need to be accurately prepared according to the detection target)

2.Chromatography materials: high performance nitrocellulose membrane (uniform pore size distribution to ensure stable chromatography flow rate), sample pad (need to be pretreated to eliminate non-specific adsorption), binding pad (to ensure uniform release of markers), absorbent pad (with enough liquid absorption capacity to maintain chromatography power), the performance of these materials directly affects the accuracy and repeatability of the detection results.

3.The provision of detection performance specifications and intended application scopes is essential for tailoring detailed solutions to clients, integrating lateral flow analysis (LFA) technology

(1) Requirement Analysis and Scheme Design

Through in-depth communication with clients, clarify the target substance for detection (such as antigen, antibody, nucleic acid, small-molecule compounds, etc.), detection scenarios (on-site testing, laboratory testing, etc.), detection performance requirements (sensitivity, specificity, detection time, etc.), and expected application scope. Based on this information, combined with the technical characteristics of lateral flow analysis, tailor a detailed development scheme for clients, including the selection of detection principles, design of reagent formulations, planning of detection processes, etc.

(2) Reagent and Material Preparation

Antibody/antigen preparation: Use hybridoma technology, genetic engineering technology, etc., to prepare highly specific and high-affinity monoclonal antibodies or polyclonal antibodies, as well as recombinant expression and purification of target antigens to ensure the quality and performance of reagents.

Marker preparation: Select appropriate markers such as colloidal gold, fluorescent microspheres, quantum dots, etc., according to detection requirements. Perform surface modification and optimization on markers to improve their binding efficiency and stability with antibodies/antigens.

Chromatographic material preparation: Carefully select high-quality chromatographic materials such as nitrocellulose membrane, sample pad, conjugate pad, absorbent pad, etc., and carry out pretreatment and performance testing to ensure that the materials meet detection requirements.

(3) Strip Assembly

Assemble the sample pad, conjugate pad, nitrocellulose membrane, absorbent pad, etc., onto the base plate in sequence according to design specifications. Cut the assembled plate into strips of specified width using precision cutting equipment. 

(4) Testing Types

Sensitivity test: Use serially diluted standards to detect the strip, determine the lowest detection limit (LOD), and evaluate the strip¡¯s ability to detect low-concentration target substances.

Specificity test: Use substances structurally similar to the target (such as cross-antigens, interfering substances, etc.) to detect the strip, investigate the specificity of the strip, and ensure it is not interfered with by other substances.

Accuracy test: Compare the detection results of the strip with reference methods (such as ELISA, PCR, etc.), calculate the coincidence rate, and evaluate the accuracy of the strip¡¯s detection results.

Precision test: Repeat the detection of the same standard on strips prepared at different times and batches, calculate the coefficient of variation (CV) of the detection results, and evaluate the precision and stability of the strip.

Stability test: Place the strip under different temperature and humidity conditions for a certain period, regularly detect its performance, investigate the stability of the strip, and determine its shelf life.

Application

1. PCR product detection

2.Food Safety Detection  

3. Rapid water quality testing

4. Disease biomarker screening

5. Protein/antibody detection

6. Genetic material (DNA/RNA) screening

7.Binding, discovery, and screening of oligonucleotides and aptamers

Support the development of multiple markers and adaptability to multiple sample types

Simplify the development process and enable rapid testing

Optimized buffer solution and membrane experimental conditions

Customized marker nanoparticle coupling

Service Process

After both parties reach an agreement through communication, the experimental plan is determined, the service requirements are set - the contract is signed - the advance payment is made - the experiment begins - the results are delivered