Lateral flow assay device used for detecting TSH level in biological sample for diagnosing hypothyroidism, comprises conjugated gold nanorods, conjugate release pad having a detecting complex having covalently-functionalized gold nanorod conjugates on membrane, adsorbent pad, test line, control line
	CHAUDHARI B P; VARMA S S
	2023-09-01
专利权人	COUNCIL SCI & IND RES SOUTH AFRICA (COUL-C)
申请日期	2023-09-01
专利号	IN202311059067-A
成果简介	NOVELTY - Lateral flow assay (LFA) device comprises: a conjugated gold nanorods, where LFA comprising sample pad; a conjugate release pad comprising a detecting complex comprising covalently-functionalized gold nanorod conjugates on the membrane; a membrane pad; an adsorbent pad; test line; and control line; where detecting complex comprises thyroid stimulating hormone (TSH) antibody conjugated to gold nanorods by covalent bonding, and gold nanorods are surface functionalized with 11-mercaptoundecanoic acid (11-MUA). The limit of detection (LOD) of the LFA is 0.3 µ international unit (IU)/ml. USE - Lateral flow assay device or kit used for detecting TSH levels in a biological sample (claimed), for diagnosing the disease, e.g., hyperthyroidism and hypothyroidism. ADVANTAGE - The LFA device having GNRs, that are highly stable and sensitive for TSH detection. DETAILED DESCRIPTION - INDEPENDENT CLAIMS are included for: 1. a method for detecting TSH levels in a biological sample using a lateral flow assay device, which involves: (a) providing a lateral flow assay device comprising covalently-functionalized gold nanorod conjugates, where gold nanorods are surface functionalized with 11-MUA and conjugated with TSH antibody; (b) applying a biological sample to the lateral flow assay device; (c) allowing the sample to migrate through the device and interact with the TSH antibody conjugated gold nanorods; and (d) detecting the presence or concentration of TSH in the sample based on the visual or instrumental detection of the gold nanorod conjugates in the device; 2. a method for producing the gold nanorods for use in the LFA device, which involves: (a) preparing gold seed particles via seed-mediated growth method by mixing an aqueous solution of chloroauric acid (HAuCl4) as a gold precursor, cetyltrimethylammonium bromide (CTAB), and ice-cold sodium borohydride (NaBH4) to obtain gold seed particles; and (b) adding gold seed particles to a growth solution containing HAuCl4as a gold precursor, CTAB as a surfactant as well as a growth directional agent and silver nitrate (AgNO3) as a growth directional agent and an ascorbic acid as a reducing agent to obtain gold nanorods; and (c) separating gold nanorods by centrifugation; 3. a method for producing surface functionalizing gold nanorods using 11-MUA, which involves: (i) providing gold nanorods by the method and 11-MUA; (ii) mixing gold nanorods and 11-MUA, followed by stirring at 25-35℃ to obtain 11-MUA modified gold nanorods; and (iii) separating 11-MUA modified gold nanorods by centrifugation at 8000 rotation/minute (rpm) for 20 minutes, 2 times, followed by dispersing in borate buffer; 4. a method for conjugating TSH antibody to surface functionalized gold nanorods, which involves: (I) providing 11-MUA modified gold nanorods by the method; (II) mixing 11-MUA modified gold nanorods and coupling reaction agents 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS), followed by gentle vortexing; (III) adding anti-TSH antibody, followed incubation for 1 hour at 4℃ to obtain Mouse Monoclonal TSH antibody (10-2427)-conjugated gold nanorods (antibody-gold nanorods conjugate); and (IV) separating Mouse Monoclonal TSH antibody (10-2427) conjugated gold nanorods by centrifugation, followed by dispersing in borate buffer; and 5. a kit for detecting TSH levels in a sample, which comprises: (i) a sample pad, (ii) a conjugate pad with immobilized TSH antibody-conjugated gold nanorods, (iii) a nitrocellulose membrane with on test line and control line, (iv) an adsorbent pad, (v) a capture antibody for test line, (vi) a secondary antibody to detection antibody for control line, and (vii) an antigen/sample, where components of the kit/strip are usually fixed to an inert backing material.
IPC 分类号	B01J-023/52 ; B82Y-030/00 ; G01N-033/543 ; G01N-033/569
国家	印度
专业领域	医药卫生
语种	英语
成果类型	专利
文献类型	科技成果
条目标识符	http://119.78.100.226:8889/handle/3KE4DYBR/20568
专题	中国科学院新疆生态与地理研究所
作者单位	COUNCIL SCI & IND RES SOUTH AFRICA (COUL-C)
推荐引用方式 GB/T 7714	CHAUDHARI B P,VARMA S S. Lateral flow assay device used for detecting TSH level in biological sample for diagnosing hypothyroidism, comprises conjugated gold nanorods, conjugate release pad having a detecting complex having covalently-functionalized gold nanorod conjugates on membrane, adsorbent pad, test line, control line. IN202311059067-A[P]. 2023.

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