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Evaluation of adjuvanted vaccines

A vaccine and immune activity technology, applied in the field of adjuvant vaccine evaluation, can solve the problems that are not yet fully understood and the impact of the reaction is complex

Inactive Publication Date: 2010-08-25
ALK ABELLO SA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the effect of a solid support on the reaction between antigen-specific IgE and antigen bound to the solid support is complex and not fully understood
Thus, to date, it has been considered impossible to measure in vitro the immunological activity of off-the-shelf solid-phase carrier vaccines containing molecular antigens, including allergenic activity and potential to induce allergic reactions, or at least to measure it accurately

Method used

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  • Evaluation of adjuvanted vaccines
  • Evaluation of adjuvanted vaccines
  • Evaluation of adjuvanted vaccines

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0145] Preparation of Allergen Vaccine with Aluminum Gel Adjuvant

[0146] Lyophilized allergens are dissolved in aqueous buffer and diluted to the desired concentration. Under stirring, "aluminum glue" (1, 3%) was added to the resulting allergen solution, followed by sterile water. The resulting solution was allowed to stand until the next day, and then the buffer was slowly added with stirring to produce the final allergen aluminum hydroxide gel.

Embodiment 1

[0149] IgE Inhibition Test on Allergens in Solution and Adsorbed on Aluminum Hydroxide Gel Adjuvant

[0150] method

[0151] On the ADVIA centaur device, IgE inhibition tests were performed. Serial dilutions (performed with TECAN (P-05-07F294)) of the inhibitor (antigen in solution or antigen-gel adjuvant vaccine) were mixed with a fixed amount of biotinylated antigen and then mixed with a solid-phase adsorbed IgE was incubated together. The amount of biotinylated allergen bound to the solid phase was estimated from the light emitted after incubation with acridinium ester-labeled streptavidin. Raw data were processed in Excel and transferred to GraphPad Prism v.4.0 for final analysis (curve fitting, graphing and statistical comparison). Fit the data to a 4-parameter logarithmic function (Equation 1):

[0152] Y = B + T - B 1 ...

Embodiment 2

[0161] Histamine Release Test of Allergen Adsorbed on Aluminum Hydroxide Gel Adjuvant

[0162] method

[0163] Histamine is measured in an ELISA-based method on the basis of competition between the histamine to be tested and its enzyme conjugate, which is used as a substrate for coating Antibody-bound tracers on microwells. The monoamine histamine is too small to completely occupy the binding site on the antibody. Accordingly, high affinity monoclonal antibodies against modified histamines have been obtained. The histamine in the sample must be derivatized in the same manner as the histamine of the conjugate. This can be easily and reproducibly achieved using acylating agents at slightly basic pH. Acylated histamine and the histamine-alkaline phosphatase conjugate in the sample compete for binding to a limited number of antibody sites when added to the microtiter wells. After incubation, the wells are washed to remove unbound components. Bound enzyme activity was then me...

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Abstract

The invention relates to an in vitro method of evaluating the immunological activity of a vaccine preparation in the form of a mixture of a molecular antigen and a carrier, wherein the mixture comprises a liquid phase and a solid phase, to which at least a part of the antigen is attached, the method comprising the steps of i) subjecting the vaccine to one or more measurements selected from the group consisting of: 1) the immunological activity of the mixture, 2) the immunological activity of antigen in the liquid phase, 3) the immunological activity of antigen in the solid phase, 4) the immunological activity of antigen in the liquid phase upon a treatment of the mixture to displace the antigen from the solid phase, and 5) the immunoligical activity of antigen in the solid phase upon a treatment of the mixture to displace the antigen from the solid phase, wherein the immunological activity measurement is selected from the group consisting of antibody binding capacity using an immunoassay employing an antigen-specific antibody bound to an antibody solid phase, b) ability to activate effector cells and c) potential for inducing anaphylaxis; and ii) using the measurement results.

Description

technical field [0001] The present invention relates to a method for evaluating in vitro the immunological activity of a vaccine preparation in the form of a mixture of an antigen and a solid carrier, said immunological activity including allergenic activity and potential to induce allergic reactions, in particular the potential to induce allergic reactions, wherein the mixture A liquid phase and a solid phase to which at least a portion of the antigen is attached are comprised. Background of the invention [0002] By mixing an aqueous solution of the antigen and a solid phase carrier, such as aluminum hydroxide gel, to produce a mixture in which at least a part of the antigen is adsorbed on the solid phase and part or none of the antigen is in the liquid phase, it can be prepared for e.g. subcutaneous injection. vaccine. The solid support can be used as an adjuvant, ie it will boost the immune response to the antigen, although the mechanism of boosting is not always fully ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/551G01N33/50G01N33/68A61K39/00A61K39/39
CPCA61K2039/55505G01N33/551G01N33/505A61K39/39G01N33/50G01N33/68G01N33/53
Inventor P·A·乌尔特赞G·伦德H·H·雅各比H-H·伊普森
Owner ALK ABELLO SA
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