Serum peptide mass spectrometry for early warning of tumor 1. Collection of basic information of the inspected person: 2. Preparation and preservation of serum: 3. Purify the serum with magnetic beads: 4. Mass spectrometric analysis of purified serum samples 5. Blind test of unknown samples Spectionmycin Lincomycin Soluble Powder Spectinomycin Hydrochloride And Lincomycin,Hydrochloride Soluble Powder,Spectinomycin Hydrochloride Livestock,Spectinomycin Hydrochloride Swine Shandong Shengli Bioengineering Co., Ltd , https://www.shenglipharm.com
1.1 Basic information of the inspected person, including age, gender, height, weight, occupation, ethnicity, eating habits, etc.
1.2 The genetic history of the family of the inspected, past illness and treatment history, allergies, etc.
1.3 The clinical manifestations of the examinee, including the performance of various physical symptoms, such as fever, pain, etc.
1.4 Clinical imaging findings of the examinee, such as nuclear magnetic, CT and PET.
1.5 Clinical examination results such as pathological sections, biopsy results, cytological observations, and tumor marker results of the examinee.
2.1 The inspected person fasted in the morning and took blood in the morning.
2.2 Using a 10-20 ml disposable syringe, 5 ml of blood was taken from the venous blood vessels of the upper arm of the examinee, and after taking out, put 10 ml of polypropylene plastic tube, and let it stand horizontally for 30 minutes at room temperature (room temperature was 25 ° C).
2.3 Centrifuge at 1400 g for 10 minutes at room temperature in a small angle centrifuge.
2.4 Remove the above serum, avoid taking out the lower layer of plasma, dispense the serum into 100 μl of each tube, and place it in a plastic tube of 200 μl of polypropylene.
2.5 A plastic tube containing 200 μl of polypropylene in serum was immediately stored at -80 °C. Can be stored for 6 months.
3.1 Thoroughly vortex the magnetic beads for at least 1 min to obtain a uniform magnetic bead suspension.
3.2 Transfer 10 μl of binding solution and 5 μl of sample (eg human serum) (containing up to 500 pmol of protein) into a standard thin-walled polypropylene PCR tube. Add 5 μl of the mixed magnetic beads, pipette up and down 5 times, and carefully mix the liquid. Wait 1 min.
3.3 Place the PCR tube in a magnetic bead separator (MBS) and wait for 20 sec to separate the beads from the supernatant. Pay attention to the movement of the magnetic beads inside the tube.
3.4 Carefully remove the supernatant with a pipette. Avoid contact between the tip and the bead, be careful not to move the bead.
3.5 Transfer the PCR tube out of the magnetic isolator.
3.6 For washing, add 100 μl of lotion.
3.7 Place the PCR tube into the magnetic isolator and move the tube back and forth 20 times between adjacent wells to wash the beads.
3.8 Wait 20 sec to collect the magnetic beads on the tube wall and carefully remove the supernatant with a gun.
3.9 Repeat steps 6-8 twice.
3.10 Add 5 μl of acetonitrile (50%) to the beads and mix thoroughly. Wait 1 min. (Caution: Acetonitrile is volatile and can evaporate quickly!).
3.11 Place the tube in the magnetic isolator and wait for 30 sec to separate the magnetic beads from the eluate and leave on the tube wall.
2.12 Finally, the eluate containing the purified polypeptide/protein was transferred to a new 0.2 ml thin-walled polypropylene PCR tube.
4.1 Add 1 μl of eluate to a new 0.2 ml thin-walled polypropylene PCR tube, add 10 μl of matrix, mix, add 1 μl on an Anchorchip target plate, and dry in air for about 5-10 minutes.
4.2 Adjust the parameters of the mass spectrometer according to the instructions, calibrate with the external standard, and then analyze the sample.
4.3 The software acquires the data, processes it, and obtains a mass spectrum for each sample.
4.4 Software analysis, establish a standard spectrum of the disease group and the normal control group, and establish a characteristic serum polypeptide profile of the disease group according to the differential protein peak.
5.1 Take the serum of the population at high risk or census, collect the data according to the above methods, collect, store, purify and analyze the serum.
5.2 Software analysis and statistics to determine whether it belongs to the disease or health.
5.3 Compare with clinical gold standards such as pathology and cytology.
5.4 Determine the blindness test accuracy of unknown samples based on statistical results.