Application of urine immunoglobulin kappa chain C region protein and polypeptide fragments thereof in lung adenocarcinoma

Abstract

The invention provides application of urine immunoglobulin kappa chain C region protein (Immunoglobulin Kappa Chain C Region, IGKC) and polypeptide fragments thereof in lung adenocarcinoma, and in particular relates to application of the urine immunoglobulin kappa chain C region protein and the polypeptide fragments thereof in preparation of a preparation for detecting and auxiliary diagnosis of the lung adenocarcinoma. Researches confirm that compared with a normal control and a lung adenocarcinoma patient group, specific expression of the urine immunoglobulin kappa chain C region protein inurine of a lung adenocarcinoma patient is up regulated, and the urine immunoglobulin kappa chain C region protein can be used for detection and auxiliary diagnosis of the lung adenocarcinoma. The advantages of non-invasive acquisition, large-scale repeated sampling, and convenient preservation of a urine specimen are played, and the urine specimen is used to detect the urine immunoglobulin kappa chain C region protein and the polypeptide fragments thereof.

Description

technical field

[0001] The present invention relates to the new application of urinary immunoglobulin kappa chain C region protein and its polypeptide fragment, in particular to the application of urinary immunoglobulin kappa chain C region protein and its polypeptide fragment in the diagnosis and treatment of lung adenocarcinoma. Background technique

[0002] Lung cancer is one of the tumors with the highest morbidity and mortality worldwide. The latest WHO statistics show that 1.59 million patients die of lung cancer every year worldwide. Lung cancer can be divided into small cell lung cancer (SCLC) and non-small lung cancer (NSCLC) according to pathological types, and NSCLC is further divided into squamous cell carcinoma, adenocarcinoma, large cell carcinoma, and sarcomatoid carcinoma. Wait. With the deepening of lung cancer research, NSCLC is divided into squamous cell carcinoma and non-squamous non-small cell lung cancer according to the existence of driving genes, an...

Examples

Embodiment 1

[0019] Example 1 Urine Specimen Collection and Processing

[0020] Randomly cleaned midstream urine samples were collected from 34 patients with lung adenocarcinoma (Beijing Shijitan Hospital Affiliated to Capital Medical University), centrifuged within 2 hours (1500rpm, 5min), and the supernatant was retained. Store in a -80°C refrigerator after aliquoting. The normal control group consisted of 36 cases (Physical Examination Center, Beijing Shijitan Hospital Affiliated to Capital Medical University). For details, please refer to the following table 1:

[0021] Table 1. Clinical data of the 2 groups

[0022]

Embodiment 2

[0023] Example 2 Magnetic Bead Purification and Isolation of Peptides from Urine Specimens

[0024] Take out the urine sample from the -80°C refrigerator, rethaw at 4°C, centrifuge (3000rpm, 10min), and take the supernatant for later use. Equilibrate Weak Cationic Magnetic Beads (MB-WCX) at room temperature and mix the magnetic bead suspension by hand. Add 10ul of MB-WCX and 10ul of magnetic bead binding buffer into the sample tube, pipette the sample gun up and down to mix well to avoid foaming. Add 5 ul of urine supernatant to the sample tube, mix well and let stand on the magnetic stand for 1 minute to separate the magnetic beads from the suspended liquid. Use a sampling gun to remove the suspended clear liquid, and the tip of the gun should avoid contact with the magnetic beads to avoid absorbing the magnetic beads. Add 100ul of magnetic bead washing buffer into the sample tube, mix well, and then place the sample tube on the magnetic stand for 1 minute, the magnetic ...

Embodiment 3

[0025] Example 3 Spot Targeting and Peptide Spectrum Generation of Urine Specimens

[0026] After calibrating the instrument with a standard, mix 1 μl of eluate with 10 μl of matrix (0.3% α-cyano-4-hydroxycinnamic acid, HCCA), and take 1 μl to spot on the Anchorchip (Autoflex MALDI TOF, Bruker-Dalton) target plate and dry at room temperature. The sample is ionized by nitrogen laser irradiation and then subjected to mass spectrometry analysis, collecting data in the range of 1000-10000 Da, and obtaining a mass spectrogram composed of protein peaks with different mass-to-charge ratios. For each MALDI crystallization point, a total of 400 laser irradiations (50 times for each crystallization point at 8 different positions) were irradiated, and the average value represented one sample, so as to obtain the peptide maps of all samples. The mass spectrograms of the normal control group and lung adenocarcinoma group were analyzed by ClinProTools2.1 analysis software, and the diffe...

technical field

[0001] The present invention relates to the new application of urinary immunoglobulin kappa chain C region protein and its polypeptide fragment, in particular to the application of urinary immunoglobulin kappa chain C region protein and its polypeptide fragment in the diagnosis and treatment of lung adenocarcinoma. Background technique

[0002] Lung cancer is one of the tumors with the highest morbidity and mortality worldwide. The latest WHO statistics show that 1.59 million patients die of lung cancer every year worldwide. Lung cancer can be divided into small cell lung cancer (SCLC) and non-small lung cancer (NSCLC) according to pathological types, and NSCLC is further divided into squamous cell carcinoma, adenocarcinoma, large cell carcinoma, and sarcomatoid carcinoma. Wait. With the deepening of lung cancer research, NSCLC is divided into squamous cell carcinoma and non-squamous non-small cell lung cancer according to the existence of driving genes, an...

Examples

Embodiment 1

[0019] Example 1 Urine Specimen Collection and Processing

[0020] Randomly cleaned midstream urine samples were collected from 34 patients with lung adenocarcinoma (Beijing Shijitan Hospital Affiliated to Capital Medical University), centrifuged within 2 hours (1500rpm, 5min), and the supernatant was retained. Store in a -80°C refrigerator after aliquoting. The normal control group consisted of 36 cases (Physical Examination Center, Beijing Shijitan Hospital Affiliated to Capital Medical University). For details, please refer to the following table 1:

[0021] Table 1. Clinical data of the 2 groups

[0022]

Embodiment 2

[0023] Example 2 Magnetic Bead Purification and Isolation of Peptides from Urine Specimens

[0024] Take out the urine sample from the -80°C refrigerator, rethaw at 4°C, centrifuge (3000rpm, 10min), and take the supernatant for later use. Equilibrate Weak Cationic Magnetic Beads (MB-WCX) at room temperature and mix the magnetic bead suspension by hand. Add 10ul of MB-WCX and 10ul of magnetic bead binding buffer into the sample tube, pipette the sample gun up and down to mix well to avoid foaming. Add 5 ul of urine supernatant to the sample tube, mix well and let stand on the magnetic stand for 1 minute to separate the magnetic beads from the suspended liquid. Use a sampling gun to remove the suspended clear liquid, and the tip of the gun should avoid contact with the magnetic beads to avoid absorbing the magnetic beads. Add 100ul of magnetic bead washing buffer into the sample tube, mix well, and then place the sample tube on the magnetic stand for 1 minute, the magnetic ...

Embodiment 3

[0025] Example 3 Spot Targeting and Peptide Spectrum Generation of Urine Specimens

[0026] After calibrating the instrument with a standard, mix 1 μl of eluate with 10 μl of matrix (0.3% α-cyano-4-hydroxycinnamic acid, HCCA), and take 1 μl to spot on the Anchorchip (Autoflex MALDI TOF, Bruker-Dalton) target plate and dry at room temperature. The sample is ionized by nitrogen laser irradiation and then subjected to mass spectrometry analysis, collecting data in the range of 1000-10000 Da, and obtaining a mass spectrogram composed of protein peaks with different mass-to-charge ratios. For each MALDI crystallization point, a total of 400 laser irradiations (50 times for each crystallization point at 8 different positions) were irradiated, and the average value represented one sample, so as to obtain the peptide maps of all samples. The mass spectrograms of the normal control group and lung adenocarcinoma group were analyzed by ClinProTools2.1 analysis software, and the diffe...

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