Customised Development: Lateral Flow Tests

  • In developing a lateral flow test for an antigen, the most important and difficult requirement to fulfill is obtaining a matching pair of monoclonal antibodies; one antibody is for coating and the other conjugates to colloidal gold.  Only with an optimal match will the results of the test have the required specificity, sensitivity, and cross-reactivity.  Moreover, the precise environment and conditions for the antibody pair needs to be determined since its activity may differ between various environments.

    For the lateral flow test development process, the client may provide us with the monoclonal antibodies, or may contract us to develop the monoclonal antibodies by providing us with the antigen.  Please see our Monoclonal Antibodies page for further details.

    The precise conditions for coating, conjugating, washing, and blocking fluctuates greatly between different tests.  Likewise, the type of materials and buffers used in the sample pad and the assembly of the rapid test can also vary.  Due to the variability and complexity of this process, pricing for the development of rapid tests differ for each individual case. Please contact us for more information.

  • Process Outline

    Strategy Planning:
    Firstly, our senior scientists will meet with the client to discuss the possibilities and limitations regarding the test the client wishes to develop. Thereafter, a detailed proposal of the developmental plans will then be presented for the client’s approval. Upon approval, scientists and technicians will be assigned to the project. Additionally, a confidentiality agreement will be signed at this point.

    Selecting Positive Antibodies:
    The rapid test development process requires approximately 150 different clones that tested positive (by ELISA) for antibody activity. The client has an option to either provide us with the selected antibodies (purified) or to have us develop and select the hybridoma clones. Reactivity, specificity, and cross-reactivity will be characterized for each clone.

    Note: We may need more than 150 clones because some antibodies may be discarded during the development process due to undesired cross-reactivity, low reactivity, and/or low sensitivity.

    Characterizing Antibody Colloidal Gold Conjugation and NC Membrane Coating: Since the optimal procedure and conditions for conjugating and coating antibodies differ for each antibody, the antibodies will be accurately characterized before the extensive pairing process begins.

    Selecting Best Matched Pair Antibody:
    Extensive comparison studies will take place by pairing all the antibodies in different combinations (as the conjugate, or as the coating antibody, with about 5000 pairs in total for trial). Two pairs that meet specificity, sensitivity, and cross-reactivity requirements will be selected.

    Note: It is entirely possible that no matching pairs will be found. In this case, cell fusion will need to be repeated, 150 new positive clones will be selected, and the selection and characterization process is to be repeated. Although this is extremely rare, the client should be financially prepared for it.

    Developing the Rapid Test:
    From the two pairs of antibodies, rapid tests will be developed with different blocking and washing methods, buffers, support membranes, and sample pads to optimize the reaction conditions on the test strip.

    Finalizing Optimal Conditions:
    Advanced technology used for the production of lateral flow tests is key to its quality. Since Artron BioResearch already possesses advanced technology for manufacturing test strips, finalizing the optimal conditions for your customized test will hardly be a challenge. The following are examples of steps of the production process that will be finalized/optimized:

      1. Conditions for conjugating MAb to colloidal gold
      2. Storage conditions for the conjugates
      3. Coating conditions for MAb and goat anti-mouse IgG
      4. Solutions and methods of blocking and washing NC membranes
      5. Type of supporting membrane (M2, M3, or M5)

    At the end of production optimization, two (2) prototype tests will be made from the two (2) pairs of antibodies, which will then undergo detailed field trials. If desired, this step can be client-conducted.

    Finalizing the Rapid Test:
    The better of the two prototypes will be selected based on sensitivity, specificity, and cross-reactivity requirements. The client will be given a final documentation of the entire process. All client-supplied antigens, antibodies, materials and intellectual properties will be returned to the client. All research documents compiled by Artron BioResearch for the project and all tests made are the property of the client.