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Detection method of anti-drug antibody and detection kit

A technology for detection kits and detection methods, applied in biological testing, measuring devices, material inspection products, etc., can solve problems such as inability to detect ADA, and achieve the effect of improving the detection effect

Active Publication Date: 2018-07-24
北京新艾进生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The object of the invention is to provide a sensitive anti-drug antibody detection method, overcome the defect that the existing method cannot detect ADA in the presence of high-concentration drugs (generally reaching more than 1 mg / mL); and overcome the bridging immunoassay method that can only detect Deficiency of free form of ADA and inability to detect ADA present in the sample as drug-ADA complex

Method used

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  • Detection method of anti-drug antibody and detection kit
  • Detection method of anti-drug antibody and detection kit
  • Detection method of anti-drug antibody and detection kit

Examples

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Effect test

preparation example Construction

[0028] 1. Preparation method of high-density enrichment magnetic beads

[0029] According to the amount of 50-500 micrograms of drug per milligram of amino magnetic beads, accurately weigh amino magnetic beads with a particle size of 0.2-2 microns and protein drugs for mixing, and add 1-(3 -Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) aqueous solution, calculated according to the addition of 20 micrograms of EDC per milligram of amino magnetic beads, shake the reaction at room temperature for 30 minutes, and use 0.01M pH7.5 Wash the magnetic beads 5 times with phosphate buffer to obtain high-density enriched magnetic beads, add 1% bovine serum albumin (BSA) solution to the high-density enriched magnetic beads, so that the mass concentration of the magnetic beads is 0.5-5%, and obtain The high-density enrichment magnetic bead reagent should be stored in a refrigerator at 4°C.

[0030] 2. Preparation method of magnetic beads for detection:

[0031] According to...

Embodiment 1

[0046] Anti-Drug Antibody Detection Kit for Tumor Necrosis Factor-α (TNF-α) Inhibitor Drug Adalimumab

[0047] The above-mentioned method was used to prepare an anti-drug antibody detection kit for Adalimumab. The kit includes high-density enrichment magnetic beads for Adalimumab ADA detection, magnetic beads for detection, Adalimumab labeled with luminescent groups, sample diluent, quality control and excitation solution.

[0048] The high-density enrichment magnetic bead reagent is that Adalimumab is bound to the surface of amino magnetic beads (250 μg Adalimumab / mg magnetic beads) with an average particle size of 0.5 μm by chemical coupling, and the high-density enriched magnetic bead is obtained by adding 1% BSA solution. Collect magnetic bead solution.

[0049] The magnetic bead reagent for detection is a solution containing magnetic beads for detection obtained by adding Adalimumab to the surface of amino magnetic beads with an average particle size of 1 μm (15 μg Adali...

Embodiment 2

[0056] Dosing inhibition experiment of Adalimumab anti-drug antibody kit

[0057] The kit in Example 1 was used to detect the anti-Adalimumab anti-drug antibody in the serum samples of cynomolgus monkeys. In order to evaluate the effectiveness of the method of the present invention for detecting ADA under the condition of high drug concentration, a drug addition inhibition experiment was carried out. The cynomolgus monkey serum without administration was used as a negative control, the cynomolgus monkey serum containing 250ng / mL rabbit anti-Adalimumab polyclonal antibody was used as ADA positive control, and the serum containing 250ng / mL rabbit anti-Adalimumab polyclonal antibody and 0.5mg / mL mL (sample A) or 1 mg / mL (sample B) cynomolgus monkey serum of Adalimumab was used as the sample to be tested, and the sample was pretreated and tested according to the aforementioned method. The results are shown in Table 1.

[0058] Table 1 Determination of the detection results of the...

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Abstract

The invention discloses a detection method of an anti-drug antibody and a detection kit. The detection method comprises the following steps of: pretreating a sample to be detected and detecting the pretreated sample, and after high-density enriching magnetic beads are adopted for enriching the sample to be detected, chemiluminesent immunoassay of magnetic particles is carried out on the sample. The detection method and the detection kit disclosed by the invention have the beneficial effects that by pretreatment from the high-density enriching magnetic beads, the detection for the total ADA (Adenosine Deaminase) existing in the form of drug-ADA complex in the sample can be realized, so that the influence of high-concentration drug protein in the sample can be overcome, and the drug resistance of the ADA detection method can be improved; simultaneously the detection method also has the advantages of fast and automatic analysis of a chemiluminesent immunoassay method. The detection kit comprises a high-density enriching magnetic bead reagent, a magnetic bead reagent for detection, a drug for labeling luminophores, sample diluted liquid, acidifying liquid, neutralizing liquid, a quality control product and exciting liquid. By adoption of the kit, the anti-drug antibodies can be rapidly and accurately detected.

Description

technical field [0001] The invention relates to the field of magnetic particle chemiluminescence immunodiagnosis, in particular to a detection method and a detection kit for anti-drug antibodies. Background technique [0002] Since the first recombinant protein drug was approved for marketing in 1982, more than 260 therapeutic recombinant protein drugs (including more than 70 therapeutic monoclonal antibody drugs) have been widely used in clinical practice at home and abroad, for more than 230 Treatment of indications (Nat Biotechnol, 2014.32(10):p.992-1000; Health Aff(Millwood), 2015.34(2):p.210-9). With the deepening of research on therapeutic recombinant protein drugs, more and more attention has been paid to the research on antibodies produced during the administration of related drugs (Anti-Drug Antibody, ADA) (Immunol Today, 1986.7( 12): p.367-8.; Front Immunol, 2016.7: p.21.). The production of anti-drug antibodies will affect the safety and effectiveness of therape...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/68
CPCG01N33/54326G01N33/6854G01N2333/47G01N2446/90
Inventor 贾向阳刘海涌陈健平贾鹏飞贾鹏耀
Owner 北京新艾进生物科技有限公司
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