Method for rapidly identifying thyroid papillary carcinoma lymphonodi cervicales metastasis in operation
A technology for lymph node metastasis and papillary carcinoma, which is applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of low contrast between color strips and background, inability to achieve quantitative detection of detected objects, and difficulty in realizing multiple inspections and joint inspections, etc. Achieve the effect of reducing external conditions and background, shortening hospital stay, improving detection sensitivity and result confidence
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Embodiment 1
[0045] Example 1: Screening of Human Tg Antigen Peptides
[0046] The Tg described in this application is known in the art, and the complete Tg is composed of a single polypeptide chain containing 2768 amino acids, with a molecular weight of about 660000 Daltons. Its amino acid sequence is known in the art and can be found in professional databases such as NCBI. The specific sequence is as follows:
[0047] Human Tg(1-2768):
[0048] MALVLEIFTLLASICWVSANIFEYQVDAQPLRPCELQRETAFLKQADYVPQCAED GSFQTVQCQNDGRSCWCVGANGSEVLGSRQPGRPVACLSFCQLQKQQILLSGYINSTDT SYLPQCQDSGDYAPVQCDVQQVQCWCVDAEGMEVYGTRQLGRPKRCPRSCEIRNRRLLH GVGDKSPPQCSAEGEFMPVQCKFVNTTDMMIFDLVHSYNRFPDAFVTFSSFQRRFPEVS GYCHCADSQGRELAETGLELLLDEIYDTIFAGLDLPSTFTETTLYRILQRRFLAVQSVISGRFRCPTKCEVERFTATSFGHPYVPSCRRNGDYQAVQCQTEGPCWCVDAQGKEMHGTR QQGEPPSCAEGQSCASERQQALSRLYFGTSGYFSQHDLFSSPEKRWASPRVARFATSCP PTIKELFVDSGLLRPMVEGQSQQFSVSENLLKEAIRAIFPSRGLARLALQFTTNPKRLQ QNLFGGKFLVNVGQFNLSGALGTRGTFNFSQFFQQLGLASFLNGGRQEDLAKPLSVGLD SNSSTGTPEAAKKDGTMN...
Embodiment 2
[0053] Embodiment 2: Preparation of TG antibody
[0054] The TG antigenic epitope peptides (1) and (2) obtained in Example 1 were linked to the carrier protein to prepare the antigens (1) and (2) for immunization respectively, and the animals were immunized with the obtained antigens (1) and (2), respectively, Antigen (1) is thus used to prepare specific monoclonal antibodies and polyclonal antibodies, and antigen (2) is used to prepare specific monoclonal antibodies and polyclonal antibodies.
[0055] 1. Antigen preparation: Tg peptides were linked with carrier protein BSA to prepare TG antigens. Take each 5.0 mg of the two antigenic peptides described in this application, dissolve them in 1 mL DMF, add 5 mg EDC (dissolved in 50 μL H 2 (0), after stirring for 20 min, the reaction was added dropwise to 5 mg BSA (dissolved in 1 mL of PBS buffer), immediately added 5 mg NHS, and stirred overnight at room temperature. Put the reaction solution into the treated dialysis bag, pla...
Embodiment 3
[0067] Example 3: Specific identification of human TG antibodies (1) and (2)
[0068] Detection was performed by ELISA. The ELISA plates were coated with human Tg protein, GAPDH protein, and neuron-specific enolase NSE as detection antigens, and the specific reactions of the prepared TG monoclonal antibodies (1) and (2) with different proteins were detected by ELISA , normal BALB / c mouse serum was used as negative control, and PBS solution was used as blank control.
[0069] Result: Tg monoclonal antibody (1) and (2) only react positively with Tg respectively (P / N>2.1), and are negative with GAPDH protein, neuron-specific enolase NSE reaction, illustrate and utilize the present invention Monoclonal antibodies (1) and (2) prepared from Tg epitope peptides (1) and (2) both have specificity.
[0070] Identification of polyclonal antibodies was performed using the same method as described above for identifying the specificity of monoclonal antibodies.
[0071] Result shows: Tg ...
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