CN102532298B - ABCC3 antigen polypeptide specially binding with autoantibody and application - Google Patents

Abstract

The invention provides an ABCC3 antigen polypeptide specially binding with an autoantibody. The antigen polypeptide is applied to detecting the corresponding specific autoantibody in bloods of lung cancer and esophageal cancer patients. The autoantibody can be used as a tumor marker to assess the medicament tolerance of lung cancer and esophageal cancer cells. The antigen polypeptide and the antibody thereof can be used for preparing early diagnosis reagent of tumors and developing a target medicament for treating tumors.

Description

ABCC3 antigenic peptide and application with the autoantibody specific binding

Technical field

The present invention relates to a kind of people ABCC3 proteantigen polypeptide, a kind of ABCC3 antigenic peptide of tumor drug resistance mark specifically, use this antigenic peptide and find ABCC3 albumen Autologous IgG antibody, IgA antibody, for the preparation of the exploitation of reagent and the targeted drug of predicting tumors resistance.Belong to field of immunology.

Background technology

Along with the development of combined therapy of tumour, the status of chemotherapy in treatment is more and more important, but chemotherapy but is not quite similar to the effect of clinical patients.Tumour cell is to cause the major cause of chemotherapy of tumors failure to Treated with Chemotherapeutic Drugs deposits yields cross resistance (Multidrug-resistance, MDR).According to the pertinent data statistics, the tumour patient cause of the death more than 90% is more or less all relevant with resistance, therefore MDR Mechanism Study and reversion MDR is become problem demanding prompt solution in oncotherapy.

Studies show that in a large number, the tumor associated antigen in serum can induce body to produce autoantibody, has both had tumour antigen in Serum of Cancer Patients, also has the autoantibody for this tumour antigen.Therefore, both can utilize the antibody test tumour antigen, and also can utilize Detection of antigen tumour autoantibody, to detect tumour with tumour antigen much higher but utilize the tumour autoantibody to detect the specificity of tumour and the equal Billy of susceptibility.A lot of tumor associated antigens not only exist in the tumour patient body, also exist in the normal human, therefore detect tumor associated antigen credible poor as diagnosis basis.And autoantibody can't detect or exists normal human's intensive amount is very low, if in body, autoantibody obviously increases, show to have the abnormal immune situation in body, show that in body, the related antigen level fluctuates, existence or original disease of indication disease increase the weight of.Have with the major advantage of tumour autoantibody as tumor markers: high specificity, highly sensitive, easy and simple to handle, good reproducibility, detection speed are fast, and its false positive rate is low.Level by the serum tumor autoantibody changes and can monitor the different steps of tumour, can carry out early diagnosis to tumour, is conducive to the early treatment of tumour, improves the prognosis of tumour patient.

Abc transport albumen full name be ATP in conjunction with box protein called membrane transporters (ATP-binding casssette transporters), they can mediate the transportation of multiple substrate molecule inside and outside cell.Be divided into 7 subfamilies (ABC A-G) according to the abc transport albumen of sequence homology and membrane-spanning domain topological framework, so far, there are 49 members in oneself finds the mankind ABC family, and the albumen relevant to drug transport mainly concentrates in ABCB, ABCC and these subfamilies of ABCG.The research discovery, the pumping function of abc transport albumen is relevant with the reduction of medicine gathering in tumour cell, is the major cause that tumour cell produces resistance, becomes tumour cell and avoids the important defense mechanism that chemotherapeutics is attacked.ABCC3 belongs to the abc transport superfamily protein, and this proteinoid can and utilize the various differing molecular cross-cell membrane transhipments of energy drives in conjunction with ATP.ABCC3 has 58% sequence homology by 1725 Amino acid profiles with ABCC1, and ABCC3 only limits to vincristine(VCR), Etoposide, teniposide, methotrexate to the resistance scope of cancer therapy drug.The most study relevant with resistance about ABCC3 concentrates on lung cancer.There are some researches show, ABCC3 just obviously increases in swollen neoplastic early expression amount, is an important molecular marker of tumor drug resistance.The present invention is by the antigen epitope polypeptide of a kind of ABCC3 of design, and detect in Serum of Cancer Patients autoantibody and develop corresponding reagent, predicting tumors patient's resistance, and provide reliable data for the tumour new drug research.

Summary of the invention

The invention provides a kind of and ABCC3 antigenic peptide and application the autoantibody specific binding.

The technical scheme that the present invention takes is: a kind of ABCC3 antigenic peptide of and autoantibody specific binding, its aminoacid sequence is Sequence N0.1.

The application of ABCC3 antigenic peptide of the present invention in preparation predicting tumors resistance test kit.

The present invention utilizes the linear polypeptide of a kind of ABCC3 drug-resistant protein of designed, designed, adopts specificity ABCC3 albumen autoantibody in ELISA method detection of lung cancer and esophagus cancer patient blood serum.Autoantibody improves the expression amount that shows ABCC3 drug-resistant protein in the tumour patient body and increases, primary or Secondary cases resistance may appear in the indication patient, can predict the effect of chemotherapy, instruct the clinician to avoid using the chemotherapeutics such as vincristine(VCR), Etoposide, teniposide, methotrexate as far as possible.

Advantage of the present invention and beneficial effect

(1) the present invention uses the ABCC3 antigenic peptide and specific autoantibody in lung cancer and esophagus cancer patient blood serum detected, and this reaction has high specific and high sensitivity, can be used for the early diagnosis of tumor drug resistance;

(2) detection of ABCC3 antigenic peptide autoantibody can be used for effect and the prognosis of auxiliary judgement clinical chemotherapy.

(3) detection of ABCC3 antigenic peptide autoantibody can be used for instructing the clinical chemotherapy medication.

(4) the ABCC3 antigenic peptide can be used for preparing the tumor drug resistance detection kit.

(new drug development that 5 the present invention can tumor drug resistance reversal agent provides the laboratory judgment basis.

Description of drawings

The SBI graphic representation that Fig. 1 ABCC3 is combined with serum IgG;

The SBI graphic representation that Fig. 2 ABCC3 antigenic peptide is combined with blood plasma IgA.

Embodiment

A kind of ABCC3 antigenic peptide of and autoantibody specific binding, its aminoacid sequence is Sequence N0.1.Purity〉95%, pH〉7.0

The application of ABCC3 antigenic peptide of the present invention in preparation predicting tumors resistance test kit.

The film functional domain of ABCC3 albumen is worn membrane structure and 1 extremely conservative ATP combined function territory by what 6 hydrophobic amino acids formed, its nucleus is by 2 cross-film zone (transmembrane domains, TMDs) and 2 Nucleotide form in conjunction with territory (nucleotide binding domans, NBDs) is common.TMDs forms the ligands specific binding site.(see Linton KJ. Structure and function of ABC transporters ［ J ］. Physiology (Bethesda), 2007,22(2): 122-130.).In fact the combination of antigen-antibody only occurs between the antigen binding site of antigenic determinant and antibody, and both are complete complementary on space structure and sterie configuration.Therefore antigenic determinant just can represent state and the affinity characteristic of whole albumen and antibodies.In addition, take recombinant protein as antigen, pass through the loaded down with trivial details processes such as vector construction, transfection, expression, screening, purifying, the albumen space structure is complicated, and epitope is difficult for exposing, so the poor specificity of antigen-antibody combination.In addition, the high sensitivity of ELISA method is high to the stability requirement of purification technique, and cost is expensive.

The contriver follows the linear polypeptide antigen of following principle design: 1. select the epicyte protein surf zone; 2. select not form the sequence of a-helix; 3. the peptide section at two ends is more reasonable than middle arrangement; 4. avoid active site of protein to repeat; 5. avoid the strong peptide section of homology; 6. avoid Cys and Glu in sequence as far as possible, too many Pro cannot be arranged, but there have 1-2 Pro to be beneficial to peptide chain structure to be stable, useful to producing specific antibody.In addition, this polypeptide antigen must contain the restricted epitope of human leucocyte two class antigen (HLA) systems, comprises HLA-DR, the restricted epitope of HLA-DP and HLA-DQ.These epi-positions can be identified by the HLA two class antigen systems of 90% above Chinese colony.

Biological characteristics based on above antigen principle of design and ABCC3 albumen, the present invention utilizes information biology and a plurality of Antigen Epitope Prediction simulation software, analyze and antigenicity associated parameter, simulate the cross-film zone of ABCC3 albumen, designed the linear aminoacid sequence Sequence N0.1 of the ligands specific calmodulin binding domain CaM of TMDs.Square frame is partly the position of polypeptide fragment in protein.

ATP-binding cassette, sub-family C (CFTR/MRP), member 3 [Homo sapiens]

1 mdalcgsgel gskfwdsnls vhtenpdltp cfqnsllawv pciylwvalp cyllylrhhc

61 rgyiilshls klkmvlgvll wcvswadlfy sfhglvhgra papvffvtpl vvgvtmllat

121 lliqyerlqg vqssgvliif wflcvvcaiv pf（rskillak aegeis）dpfr fttfyihfal

181 vlsalilacf rekppffsak nvdpnpypet sagflsrlff wwftkmaiyg yrhpleekdl

241 wslkeedrsq mvvqqlleaw rkqekqtarh kasaapgkna sgedevllga rprprkpsfl

301 kallatfgss flisacfkli qdllsfinpq （llsilirfis npmaps）wwgf lvaglmflcs

361 mmqslilqhy yhyifvtgvk frtgimgviy rkalvitnsv krastvgeiv nlmsvdaqrf

421 mdlapflnll wsaplqiila iyflwqnlgp svlagvafmv lliplngava vkmrafqvkq

481 mklkdsrikl mseilngikv lklyawepsf lkqvegirqg elqllrtaay lhttttftwm

541 cspflvtlit lwvyvyvdpn nvldaekafv svslfnilrl plnmlpqlis nltqasvslk

601 riqqflsqee ldpqsverkt ispgyaitih sgtftwaqdl pptlhsldiq vpkgalvavv

661 gpvgcgkssl vsallgemek legkvhmkgs vayvpqqawi qnctlqenvl fgkalnpkry

721 qqtleacall adlemlpggd qteigekgin lsggqrqrvs laravysdad ifllddplsa

781 vdshvakhif dhvigpegvl agktrvlvth gisflpqtdf iivladgqvs emgpypallq

841 rngsfanflc nyapdedqgh ledswtaleg aedkeallie dtlsnhtdlt dndpvtyvvq

901 kqfmrqlsal ssdgegqgrp vprrhlgpse kvqvteakad galtqeekaa igtvelsvfw

961 dyakavglct tlaicllyvg qsaaaiganv wlsawtndam adsrqnntsl rlgvyaalgi

1021 lqgflvmlaa mamaaggiqa arvlhqallh nkirspqsff dttpsgriln cfskdiyvvd

1081 evlapvilml lnsffnaist lvvimastpl ftvvilplav lytlvqrfya atsrqlkrle

1141 svsrspiysh fsetvtgasv iraynrsrdf eiisdtkvda nqrscypyii snrwlsigve

1201 fvgncvvlfa alfavigrss lnpglvglsv syslqvtfal nwmirmmsdl esnivaverv

1261 keysktetea pwvvegsrpp egwpprgeve frnysvryrp gldlvlrdls lhvhggekvg

1321 ivgrtgagks smtlclfril eaakgeirid glnvadiglh dlrsqltiip qdpilfsgtl

1381 rmnldpfgsy seediwwale lshlhtfvss qpagldfqcs eggenlsvgq rqlvclaral

1441 lrksrilvld eataaidlet dnliqatirt qfdtctvlti ahrlntimdy trvlvldkgv

1501 vaefdspanl iaargifygm ardagla

By above protein sequence as can be known, this linear polypeptide antigen is comprised of 30 amino-acid residues, contains altogether 6 overlapping epi-positions, can detect at least 6 kinds of monoclonal antibodies, has the specificity of height.In addition, two portions polypeptide that consists of this linearity antigen all is exposed to surface of cell membrane, is conducive to identification and the combination of antibody.Therefore, can infer its drug-resistant intensity according to antibody horizontal in the patient body, the assessment chemotherapy effect.

The ELISA method detects autoantibody

(1) enzyme plate design: each plasma sample is established people ABCC3 antigenic peptide two multiple holes, the two multiple holes of goat polypeptide contrast antigen (gAg) and negative control two multiple holes (NC).GAg antigen and human protein organize without homology, and purpose is the interference that reduces the non-specific binding reaction, the working concentration scope of gAg〉15 μ g/ml.

(2) coated: antigenic peptide is coated in 96 hole enzyme plate (COSTAR with coating buffer (pH7.0～7.4 0.01M PBS/0.1%NaN3), the U.S.), every hole 100 μ l, the ABCC3 antigenic peptide is coated with concentration 7.5～15.0 μ g/ml, the coated concentration of gAg is 15～20 μ g/ml, and 4 ℃ are spent the night.

(3) primary antibodie/blood plasma is hatched: 0.01M PBS/0.005% TWEEN-20 cleans every hole 3 times, utilize analytic liquid (0.01M PBS+1%BSA+2% sheep blood plasma) with blood plasma 1:500 dilution (surveying IgG) or 1:200 dilution (surveying IgA), every hole 100 μ l are hatched 2～3h for 25 ℃;

(4) two anti-hatching: 0.01M PBS/0.005% TWEEN-20 cleans every hole 5 times, utilize the goat anti-human igg (U.S. of analytic liquid (ditto) dilution horseradish peroxidase-labeled, Sigma) or the goat-anti people IgA (U.S. of horseradish peroxidase-labeled, Sigma), every hole adds 200 μ l, hatches 2h for 25 ℃; The goat anti-human igg ELISA working range of horseradish peroxidase-labeled: 1:30000～1:50000, the goat-anti people IgA working range of horseradish peroxidase-labeled: 1:20000～1:50000.

(5) colour developing: 0.01M PBS/0.005% TWEEN-20 cleans every hole 5 times, utilizes 3,3', and 5,5'-tetramethyl benzidine (TMB) is the substrate (Invtrogen, the U.S.) of peroxidase, and every hole adds 100 μ l, room temperature lucifuge 15～30min.

(6) detect: every hole adds 50 μ l 10%H ₂SO ₄Be reaction terminating liquid, use microplate reader (BioTeck ELx800, the U.S.) to detect the OD value in 10min, the detection wavelength is 450nm, and reference wavelength is 630nm.

Each sample of Quality Control is established two multiple holes, is averaged the OD value.OD value plastisied dispersion is judged: plastisied dispersion=OD1-OD2/OD1+OD2, and plastisied dispersion≤0.1 is effective result; Plastisied dispersion〉0.1, be null result.Getting 100 parts of Healthy Human Serum equal-volumes mixes as Quality Control blood (Quality control, QC), represent crowd's common situation, every plate is all established 2 QC blood plasma holes, with the stability of the OD value Deflection level result of determination in QC blood plasma hole, all batches of SD/ QC hole, batch variation CV=all batches QC hole OD average＜20%.SD/ each plate QC every day hole, variation within batch CV=each plate QC every day hole average＜10%.

Data analysis adopts SPSS17.0 for windows to carry out statistical analysis.Adopt specific combination index (Specific binding index, SBI) to judge the combination degree of ABCC3 antigenic peptide and blood plasma autoantibody, SBI=ABCC3 OD value – NC OD/ gAg OD value – NC OD, NC is the negative control of each sample.The ROC curve is that take True Positive Rate (sensitivity) as ordinate zou, false positive rate (1-specific degree) is the curve that X-coordinate is drawn according to a series of two different mode classifications (cut off value or decision threshold).ROC area under a curve value is between 1.0 and 0.5.In the situation that AUC〉0.5, AUC illustrates that more close to 1 diagnosis accuracy is better.The ROC curve combines sensitivity and specificity with graphic technique, can accurately reflect the relation of certain analytical procedure specificity and susceptibility, is the comprehensive representative of test accuracy.Analyse-it for Microsoft Excel Software on Drawing ROC curve is adopted in this invention, and area under calculated curve (AUC) is judged to the positive with Healthy People SBI mean value+2SD, judges sensitivity and specific degree; Carry out sum of ranks ( Z）Check, check one class mistake level is a=0.05.

The process that embodiment 1 ABCC3 antigenic peptide is combined with plasma specific IgG

As shown in Figure 1, during ABCC3 concentration 5～10 μ g/ml, along with the increase of concentration, the SBI value descends gradually, and when ABCC3 antigenic peptide concentration 10～20 μ g/ml, along with the increase of concentration, the SBI value rises gradually and tends to be steady.This SBI binding curve shows, when the ABCC3 antigenic peptide is 5 μ g/ml(0.5 μ g/well) low concentration the time, be not paved with at the bottom of the plate of 96 hole enzyme plates, cause nonspecific reaction high, so this moment, the SBI value was higher, be false positive results; Along with ABCC3 antigenic peptide Enrichment, at the bottom of antigen is paved with whole plate gradually, its blocking effect manifests, nonspecific reaction reduces gradually, during to 10 μ g/ml nonspecific reaction is minimum, and this moment, the ABCC3 antigenic peptide began to occur with the specific binding of IgG antibody, and along with the enhancing gradually of increasing of antigen concentration, during to 15 μ g/ml, bonding force is the strongest, tends to be steady afterwards.This SBI binding curve has fully represented the specific binding reaction process of Autologous IgG antibody in ABCC3 antigenic peptide and blood plasma.

The process that embodiment 2 ABCC3 antigenic peptides are combined with plasma specific IgA

As shown in Figure 2, during ABCC3 antigenic peptide concentration 5～10 μ g/ml, along with the increase of concentration, the SBI value descends gradually, and when ABCC3 antigenic peptide concentration 10～20 μ g/ml, along with the increase of concentration, the SBI value rises gradually and tends to be steady.This SBI binding curve shows, when the ABCC3 antigenic peptide is 5 μ g/ml(0.5 μ g/well) low concentration the time, be not paved with at the bottom of the plate of 96 hole enzyme plates, cause nonspecific reaction high, so this moment, the SBI value was higher, be false positive results; Along with ABCC3 antigenic peptide Enrichment, at the bottom of antigen is paved with whole plate gradually, its blocking effect manifests, along with antigen concentration increases, non-specific binding reduces gradually, and during to 10 μ g/ml, nonspecific reaction is minimum, this moment, the specific binding of ABCC3 antigenic peptide and IgA antibody began to occur, and along with increasing gradually of antigen concentration strengthens, during to 15 μ g/ml, bonding force is the strongest, tends to be steady afterwards.This SBI binding curve has fully represented in ABCC3 antigenic peptide and serum the specific binding reaction process of self IgA antibody.

Embodiment 3

The test kit preparation

1 reagent reagent is formulated as follows:

1.1 antigen coated damping fluid

Sodium azide 0.1g

4 ℃ of preservations of PBS (0.01M, pH7.4) 100ml

1.2 0.1M PBST lavation buffer solution (100ml volume)

NaCl 8g

KCl 0.2g

Na ₂HPO ₄.12H ₂O 2.9g

KH ₂PO _{4 0.2g}

Tween-20 0.5ml

4 ℃ of preservations are diluted 10 times before use;

1.3 diluted sample analytic liquid

BSA 1.0g

100% lowlenthal serum 2ml

0.01M PBS up to 100ml-20 ℃ preservation

1.4 stop buffer

Dense H ₂SO ₄10.9ml

Distilled water 89.1ml

Up to 100ml room temperature preservation

1.5 work antigen

ABCC3 polypeptide antigen 5mg

67% acetic acid 1ml

-20 ℃ of preservations are diluted to 7.5～15 μ g/ml with coating buffer before use;

1.6 reference antigen

gAg 5mg

67% acetic acid 1ml

Be diluted to 20 μ g/ml with coating buffer before use ,-20 ℃ of preservations;

1.7 two anti-reference liquids

HRP-IgG working range: 1:30000～1:50000

HRP-IgA working range: 1:20000～1:50000

-20 ℃ of preservations

The substrate nitrite ion

3,3', 5,5'-tetramethyl benzidine (TMB), 100 μ l/well ,-4 ℃ of preservations;

2 operations

(1) coated: work antigen and reference antigen are diluted to working concentration with coating buffer, are coated in enzyme plate, and 4 ℃ are spent the night.

(2) add blood plasma (primary antibodie): enzyme plate is used lavation buffer solution and is cleaned 3 times, utilize analytic liquid with diluted plasma to suitable concn, be generally 1:200～1:500, every hole 100 μ l, 25 ℃ or incubated at room 2～3h;

(3) two anti-hatching: lavation buffer solution cleans 3～5 times, utilizes analytic liquid dilution two anti-reference liquid IgG or IgA, and every hole adds 200 μ l, 25 ℃/incubated at room 2h;

(4) colour developing: lavation buffer solution cleans 3～5 times, and every hole adds 100 μ l substrate nitrite ions, room temperature lucifuge 15～30min.

(5) detect: every hole adds 50 μ l stop buffers, and 10min detects, and wavelength is 450nm, and reference wavelength is 630nm.

Embodiment 4

The ABCC3 autoantibody of patients with lung cancer detects

1 sample collection is collected 486 parts of tumour patients and human normal plasma sample.Healthy group 219 examples, age (57.07 ± 10.36) year, male 134 examples wherein, female's 92 examples.Lung cancer group 267 examples.Healthy group with the lung cancer group in sex, age-matched, have comparability ( P0.05).The medical diagnosis on disease of this invention is according to international disease classification standard ICD-10.

2 detected results by Tab.10 as can be known, in Plasma of The Patients With Lung Cancer, the IgG antibody ROC area under curve (AUC) of ABCC3 antigenic peptide is 0.61, sensitivity is 19.1%, specific degree is 90%; The AUC of the IgA antibody of ABCC3 polypeptide antigen is 0.57, and sensitivity is 17.1%, and specific degree is 90%.The IgG antibody positive rate of being combined with the ABCC3 polypeptide antigen in Plasma of The Patients With Lung Cancer higher than the health group ( Z=4.18, P＜0.0001), the IgA Positive rate also apparently higher than the health group ( Z=2.93, P=0.0017).In the detection of IgA level, SBI is positive, and threshold value is 1.4081, and patients with lung cancer positive rate 5.99%, Healthy People positive rate are 4.57%.The SBI mean value of healthy group is (1.0233 ± 0.1924), and patients with lung cancer SBI mean value is (1.0774 ± 0.2202), patients with lung cancer apparently higher than the health group ( t=2.9488, P＜0.01), lung squamous cancer SBI mean value (1.0947 ± 0.2382), apparently higher than the health group ( t=2.8031, P＜0.01), adenocarcinoma of lung SBI mean value is (1.0646 ± 0.2056), also higher than the health group ( t=2.0089, P=0.0453).In the IgG level detection, SBI is positive, and threshold value is 1.4336,, the Healthy People positive rate is 2.74%, patients with lung cancer positive rate 8.61%, the positive rate of patients with lung cancer are 3.14 times of Healthy People.The SBI mean value of healthy group is (1.1636 ± 0.1350), and patients with lung cancer SBI value is (1.2211 ± 0.1552), apparently higher than the health group ( t=4.3673, P＜0.01), in different pathological types, lung squamous cancer SBI mean value (1.2004 ± 0.1505), apparently higher than Healthy People ( T=2.2692, P=0.0239), adenocarcinoma of lung SBI mean value is (1.2365 ± 0.1573), higher than health group level ( t=4.6596, P＜0.01).Above data fully show, utilizing the designed antigenic peptide of the present invention to detect the patients with lung cancer autoantibody IgG/IgA level and the normal health group that obtain relatively has obvious significant difference.

The ABCC3 autoantibody of table 1 lung cancer detects ROC tracing analysis result

Antibody	AUC	SE	95% CI	Z	P	Sensitivity(%)	Specificity(%)
								IgG	0.61	0.027	0.56-0.66	4.18	<0.0001	19.1	90
IgA	0.57	0.026	0.52-0.62	2.93	0.0017	17.1	90

Embodiment 5

Patient with esophageal carcinoma ABCC3 autoantibody detects

1 sample 314 parts of tumour patients of source collection and human normal plasma sample.Healthy group 219 examples, age (57.07 ± 10.36) year, male 134 examples wherein, female's 92 examples.Esophagus cancer 95 examples.Healthy group with esophageal carcinoma group in sex, age-matched, have comparability ( P0.05).The medical diagnosis on disease of this invention is according to international disease classification standard ICD-10.

2 detected results by Tab.11 as can be known, in esophagus cancer patient blood serum, the ROC area under curve (AUC) of the IgG antibody of ABCC3 polypeptide antigen is 0.54, sensitivity is 14.8%, specific degree is 90%; The IgA antibody A UC of ABCC3 polypeptide antigen is 0.65, and sensitivity is 23.2%, and specific degree is 90%.In patient with esophageal carcinoma blood plasma the IgG Positive rate close with healthy group ( Z=1.12, P=0.132), but the IgA Positive rate apparently higher than the health group ( Z=4.48, P＜0.0001).In the detection of IgA level, the SBI mean value of healthy group is (1.0233 ± 0.1924), and patient with esophageal carcinoma SBI mean value is (1.1306 ± 0.2142), apparently higher than the health group ( t=4.4088, P=0.0000), SBI is positive, and threshold value is 1.4081, and the Healthy People positive rate is 4.57%, and esophageal carcinoma positive rate is 12.63%, and the patient with esophageal carcinoma positive rate is 2.76 times of Healthy People.In the IgG level detection, the SB mean value of healthy group is (1.1636 ± 0.1350), and patient with esophageal carcinoma SBI mean value is (1.1842 ± 0.1566), be on close level with the health group ( t=1.1775, P=0.2399), SBI is positive, and threshold value is 1.4336, and the Healthy People positive rate is 2.74%, and esophageal carcinoma positive rate is 4.21%.Utilize the designed antigenic peptide of the present invention to detect the patient with esophageal carcinoma autoantibody IgA level and the normal health group that obtain obvious significant difference is relatively arranged.

The ABCC3 autoantibody of table 2 esophageal carcinoma detects ROC tracing analysis result

Antibody	AUC	SE	95%CI	Z	P	Sensitivity(%)	Specificity(%)
								IgG	0.54	0.027	0.47-0.62	1.12	0.132	14.8	90
IgA	0.65	0.034	0.59-0.72	4.48	<0.0001	23.2	90

SEQUENCE LISTING

<110> Liu, Linlin

＜120〉with ABCC3 antigenic peptide and the application of autoantibody specific binding

<130> Liu2012001

<160> 1

<170> PatentIn version 3.5

<210> 1

<211> 30

<212> PRT

＜213〉people

<400> 1

Leu Leu Ser Ile Leu Ile Arg Phe Ile Ser Asn Pro Met Ala Pro Ser

1 5 10 15

Arg Ser Lys Ile Leu Leu Ala Lys Ala Glu Gly Glu Ile Ser

20 25 30

Claims (2)

1. the ABCC3 antigenic peptide with the autoantibody specific binding, is characterized in that its aminoacid sequence is SEQ ID NO.1.

2. the application in the test kit of the ABCC3 albumen autoantibody IgG of the ABCC3 antigenic peptide of as claimed in claim 1 and autoantibody specific binding in preparation detection of lung cancer or esophagus cancer patient blood serum, IgA level.

Images