Desorption composition of adsorption type vaccine containing CpG ODN and application of desorption composition

Abstract

The invention provides a desorption composition of an adsorption type vaccine containing CpG ODN, and the desorption composition is prepared from potassium phosphate, an M199 culture medium, magnesium chloride and Triton-100. On the other hand, the invention provides a kit for the adsorption type vaccine containing the CpG ODN adjuvant, and the kit comprises the desorption composition. On the other hand, the invention provides a method for detecting the efficacy of the adsorption type vaccine containing the CpG ODN adjuvant in vitro. On the other hand, the invention provides the composition, the kit, the detection method and application of the composition, the kit and the detection method in desorption and in-vitro potency detection of the adsorption type vaccine containing the CpG ODN adjuvant. The desorption composition provided by the invention fills the blank in the aspect of antigen dissociation of the adsorption type vaccine containing the CpG ODN adjuvant, provides a rapid and accurate method for determining the potency of the adsorption type vaccine containing the CpG ODN adjuvant, and has pioneering significance.

Description

technical field

[0001] The present invention relates to the determination of the efficacy of an adjuvant-containing vaccine, in particular to a desorption composition in an adsorption-type vaccine containing CpG ODN and a method for determining the efficacy of an antigen in an adsorption-type vaccine containing CpG ODN. Background technique

[0002] Adjuvant is an important component of many human vaccines. The addition of adjuvant to vaccines can enhance the body's immune response to antigens. Aluminum adjuvant is the earliest and most widely used vaccine adjuvant. The adsorption of aluminum adjuvant to antigen is affected by its own physical and chemical properties, the ionic strength of the solution and the concentration of the antigen. to enhance the immune response.

[0003] CpG ODN (CpG oligonucleotide) is a representative of new adjuvants, which can bind to Toll-like receptors on the cell membrane and induce the production of various cytokines. After binding to TLR...

Examples

Embodiment 1

[0063] This implementation provides a method for detecting antigen efficacy in a CpG ODN composite adjuvant adsorption vaccine, the specific steps are as follows:

[0064] Step 1: Preparation of desorption reagents: wherein the final molar concentration of potassium phosphate is 0.6M, the final molar concentration of magnesium chloride is 0.1M, the final volume concentration of Triton-100 is 1%, and the final volume concentration of M199 medium is 20% ;

[0065] Step 2: Desorb the CpG ODN composite adjuvant-adsorbed vaccine reference substance and the sample to be tested: Mix the reference substance and sample with the desorption reagent in a volume ratio of 1:1, incubate at 28°C for 60 minutes, and incubate every 20 minutes during the incubation process. Invert and mix once;

[0066] The obtained mixture was centrifuged, the centrifugation program was 6000rpm, the centrifugation time was 20min, and the centrifugation temperature was 4°C to obtain a supernatant;

[0067] Ste...

Embodiment 2

[0075] This implementation provides the results of antigen dissociation in different batches of CpG ODN composite adjuvant-adsorbed novel coronavirus inactivated vaccines, and examines the applicability of the dissociation method to different batches of samples.

[0076] The specific desorption and antigen potency determination methods are the same as those in Example 1.

[0077] Table 4 shows the test results of antigen potency in different batches of CpG ODN composite adjuvant-adsorbed novel coronavirus inactivated vaccines.

[0078] Table 4

[0079]

[0080] Sample 1-5 double parallel lines result as figure 2 shown.

[0081] As can be seen from Table 4, the present invention has good applicability to the CpG ODN composite adjuvant adsorption type novel coronavirus inactivated vaccine.

Embodiment 3

[0083] This example verifies the applicability of desorption of CpG ODN composite adjuvant-adsorbed new inactivated vaccines using desorption compositions with different component ratios, different desorption conditions, and different centrifugation parameters.

[0084] The theoretical antigen concentration of the sample to be detected used in this example is 13U / ml, and the sample dilution scheme is 2X, 4X, 8X, and 16X. Antigen potency test results between 0.5-2.0 are considered qualified.

[0085] Table 5 shows the desorption conditions used in each sample group and the detection results of antigen potency.

[0086] The specific antigen potency determination operation procedure is the same as that in Example 1.

[0087] table 5

[0088]

[0089] Samples 1-6 double parallel line results such as image 3 shown.

[0090] It can be seen from Table 5 that the test results are all qualified, and it is verified that the desorption composition, the desorption reagent and the ...

technical field

[0001] The present invention relates to the determination of the efficacy of an adjuvant-containing vaccine, in particular to a desorption composition in an adsorption-type vaccine containing CpG ODN and a method for determining the efficacy of an antigen in an adsorption-type vaccine containing CpG ODN. Background technique

[0002] Adjuvant is an important component of many human vaccines. The addition of adjuvant to vaccines can enhance the body's immune response to antigens. Aluminum adjuvant is the earliest and most widely used vaccine adjuvant. The adsorption of aluminum adjuvant to antigen is affected by its own physical and chemical properties, the ionic strength of the solution and the concentration of the antigen. to enhance the immune response.

[0003] CpG ODN (CpG oligonucleotide) is a representative of new adjuvants, which can bind to Toll-like receptors on the cell membrane and induce the production of various cytokines. After binding to TLR...

Examples

Embodiment 1

[0063] This implementation provides a method for detecting antigen efficacy in a CpG ODN composite adjuvant adsorption vaccine, the specific steps are as follows:

[0064] Step 1: Preparation of desorption reagents: wherein the final molar concentration of potassium phosphate is 0.6M, the final molar concentration of magnesium chloride is 0.1M, the final volume concentration of Triton-100 is 1%, and the final volume concentration of M199 medium is 20% ;

[0065] Step 2: Desorb the CpG ODN composite adjuvant-adsorbed vaccine reference substance and the sample to be tested: Mix the reference substance and sample with the desorption reagent in a volume ratio of 1:1, incubate at 28°C for 60 minutes, and incubate every 20 minutes during the incubation process. Invert and mix once;

[0066] The obtained mixture was centrifuged, the centrifugation program was 6000rpm, the centrifugation time was 20min, and the centrifugation temperature was 4°C to obtain a supernatant;

[0067] Ste...

Embodiment 2

[0075] This implementation provides the results of antigen dissociation in different batches of CpG ODN composite adjuvant-adsorbed novel coronavirus inactivated vaccines, and examines the applicability of the dissociation method to different batches of samples.

[0076] The specific desorption and antigen potency determination methods are the same as those in Example 1.

[0077] Table 4 shows the test results of antigen potency in different batches of CpG ODN composite adjuvant-adsorbed novel coronavirus inactivated vaccines.

[0078] Table 4

[0079]

[0080] Sample 1-5 double parallel lines result as figure 2 shown.

[0081] As can be seen from Table 4, the present invention has good applicability to the CpG ODN composite adjuvant adsorption type novel coronavirus inactivated vaccine.

Embodiment 3

[0083] This example verifies the applicability of desorption of CpG ODN composite adjuvant-adsorbed new inactivated vaccines using desorption compositions with different component ratios, different desorption conditions, and different centrifugation parameters.

[0084] The theoretical antigen concentration of the sample to be detected used in this example is 13U / ml, and the sample dilution scheme is 2X, 4X, 8X, and 16X. Antigen potency test results between 0.5-2.0 are considered qualified.

[0085] Table 5 shows the desorption conditions used in each sample group and the detection results of antigen potency.

[0086] The specific antigen potency determination operation procedure is the same as that in Example 1.

[0087] table 5

[0088]

[0089] Samples 1-6 double parallel line results such as image 3 shown.

[0090] It can be seen from Table 5 that the test results are all qualified, and it is verified that the desorption composition, the desorption reagent and the ...

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