JPH07508352A - Tumor marker control - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Tumor marker comparison field of invention The invention generally relates to methods for use in in VitrO diagnostic assays. , regarding human serum-based stable controls, and more specifically for tumor markers. Human resources for use in monitoring the accuracy of all in vitro diagnostic assays. For a stable control based on human serum.

Background of the invention Tumor markers are substances released into the bloodstream by tumor cells. tumor marker -can be detected in serum and other body fluids, and has been shown to treat a variety of sensitive tumors clinically. This is an action to be monitored.

The term tumor marker refers to an oncogene or abnormally expressed protein, e.g. Enzymes, hormones, and receptors that are associated with and help identify tumor types It has been expanded to include characteristics of cells or tissues, such as

Clinical oncologists are responsible for diagnosing patient conditions, estimating prognosis and monitoring treatment. To assist, the presence and/or amount of these markers in body fluids is determined.

Serum assays for tumor markers are commercially available. These serum assays Oimmunoassay, enzyme immunoassay, fluorescent immunoassay and other clinical tests It is carried out using assay systems such as bed analysis techniques.

Control and monitor the accuracy, precision, and reliability of these assay systems. It is important to ensure that the patient is receiving the correct treatment and that the assay results are medically appropriate. It is extremely important to ensure that

Several human serum-based controls are currently commercially available. The control is generally within a level representing a particular range, e.g. high, low and/or normal range. Ru. These commercially available tumor control products include Cancer from POLYλIEDcO. r Antigen Controls, NMS Pharmaceuti T-8 IARKERS from cals, Inc. QtJAL [TY C0NT R0L SERUM, and LYPI from 810-RAD (OCHEK Tu mor Marker Control is included. Currently available controls are , however, many of the tumor markers needed by clinical oncologists Lacking. Furthermore, currently available commercial controls have limited transparency, limited It has lyophilization stability and limited post-reconstitution stability. In addition, commercially available control Nowadays, there are only two levels of contrast (i.e. ``high'' and ``low''). There is. In addition, some of the concentrations of ingredients in commercial controls may be too high to be fully useful. Some are too high or too low.

Although methods have been published for purifying some tumor markers, these methods Many of the methods are laborious and require several steps. Each additional step increases the collection of tumor markers. resulting in a decrease in rate. To use as a control, tumor markers should be It does not necessarily require the high degree of purity that can be obtained with some purification methods.

Thus, we have a broader range of tumor markers and their individual useful ranges. In contrast, a tumor marker with There is a demand for it. It also provides high yields and purity, which is sufficient for this intended application. Easier purification methods for some of these tumor markers may lead to increased There is also a demand for

Summary of the invention The human-based serum or plasma controls of the invention are preferably for patient diagnosis. Contains many of the tumor markers utilized by clinical oncologists. According to the present invention, The serum or plasma control based on Moreover, the optical transparency after restoration is improved.

detailed description of the invention The human-based serum or plasma control of the present invention is comprised of human male defatted plasma or serum. The tumor marker includes a tumor marker. The tumor marker includes adrenocorticotropic hormone (ACTH), aldosterone, α-fetoprotein (AFP), β2-mic Loglobulin (82M), CA 15-3■CA125■, CA 19-90, CA 19-90 (Centocor Inc.

(registered trademark of Centocor Diagnostics, a division of Centocor Diagnostics), C A349, carcinoembryonic antigen (CEA), ferritin, gastrin, human chorionic Gonadotropin (hCG), β-hCG, γ-enolase (NSE), Prola cutin, prostatic acid phosphatase (PAP), prostate-specific antigen (PSA) , tissue polypeptide antigen (TPA), calcitonin and LD-1. good. The control should also contain a preservative system. As the preservative system, freezing It must contain a preservative that is stable both before drying and after lyophilization. stomach.

Highly sensitive to certain markers, especially proteases produced by microorganisms A preservative system is required to ensure the stability of highly sensitive enzymes after reconstitution. Assemble A combined preservative is added. A preferred preservative system is gentamicin sulfate, Loheximide and Proclin 300 (Rohm and It is a combination of 　Haas). Broclin 300, however, cannot be lyophilized. Not effective after drying; useful to control microbial growth during manufacturing process . Gentamicin sulfate and cycloheximide are used to control microbial growth after reconstitution. used for Primarily due to the hazardous nature of azide's explosive nature, sodium azide is lium is not used.

The stability of the lyophilized control should be at least about 1 year, preferably at least It is also about 3 years. Post-reconstitution stability for most components requires at least 7 days and is preferred. Preferably at least about 14 days.

The base consists of human serum or plasma to preserve the stability of the PAP marker. In addition, virtually all donors must come from male donors. virtually all male In the presence of serum or plasma, PAP is very stable. female serum and Plasma may contain antibodies against this enzyme marker. There is the antibody would effectively exclude PAP from the control solution. If a woman's blood whether those antibodies are successfully removed from the serum or plasma or their effects are eliminated. If so, the serum or plasma obtained could be used as a base material.

The amount of lipids in human serum or plasma must be reduced. The lipid content is Treatment of serum or plasma with silica microparticles or dextran sulfate or other known can be reduced by a method. The methods used to reduce lipids The content of cholesterol and triglycerides in the serum or plasma of Each must be guaranteed to be less than 20rn g/dL. lipid content There are at least three reasons why this should be reduced.

First, the stability of added tumor markers that are susceptible to denaturation or oxidation may be affected by lipid content. Increases when decreased. This means that when lipids break down, some of the markers This is because it produces oxidation by-products that can impair the stability of the product. Furthermore, the decomposition of lipids results in cloudy solution.

Second, the reduction in lipids aids the restorative process. The lyophilized control is the one in which lipids have been removed. If the liquid is added, it will be restored immediately when the liquid is added.

If serum or plasma containing normal amounts of lipids is used, the reconstitution of the lyophilized control There will be a delay of approximately 15 to 30 minutes.

Third, high levels of lipids can cause interference with the measurement of some tumor markers. becomes. Thus, reduced levels of lipids lead to more accurate assay results. .

The serum or plasma used as the base for the control is free of tumor marker addition. The tumor marker to be added must be assayed beforehand.

Table 1 is a classification of different types of tumor markers added to the base material.

Table ■ lists tumor markers and the cancer types that are usually associated with the marker. Ru. Table ■ lists several sources of tumor markers.

The tumor marker added to the base material is not relatively pure, i.e. not cross-contaminated with other markers or contaminated with interfering substances. Must. It is best to use a source of tumor markers that is naturally human-like. However, many tumors of human origin produce more than one marker. It has been found that. The addition of this ingredient to the base material allows for accurate detection of each tumor marker. making it difficult to prescribe controls with volume. In some cases, tumor markers may end up being added in too high an amount to be used for low or normal control levels. It can become. Therefore, to avoid this problem, many tumor markers have to be Must be purified to remove differential contamination. In addition, added to the base material Tumor markers to be tested should be measured for cross-contamination and the presence of known interfering substances. must be assayed to determine the

82M can be purified from urine collected from patients with renal impairment. grain The urine is diafiltered into an appropriate buffer to an appropriate concentration and concentrated. 82 M (which is a protein) has an approximate molecular weight of about 11.000 Daltons . Therefore, it is suitable for size exclusion chromatography such as gel filtration chromatography. Can be refined using fees. A preferred gel abrasive is Ultragel A CA 54 or equivalent. Both portions containing 28M are pooled and preferably or at least about 1 g/dLl concentration and then subjecting the purified product to electrophoresis. can be quantified using known methods. In addition, 82M is a commercially available immuno Tested by assay. 82M is about 2-8°C or below -20°C Stable when frozen and stored at low temperatures. 82M obtained is 82M useful Removes impurities consisting of up to approximately 70% immunoglobulin without affecting sex. It can contain.

CA125 is a marker specific to ovarian cancer. This marker is associated with ovarian cancer can be found in ascitic fluid taken from patients with

The ascites contains two markers, CAl25 and TPA. Level of contamination by TPA The bell is very high, therefore add the exact amount of each CA125 and TPA. , those markers must be separated. Both of these markers and also leak into the serum during tumor growth, and due to the similarity of these markers Their separation is difficult. TP’A is a phosphate buffer solution at approximately physiological pH. Among them, Phenyl Sepharose, a hydrophobic interaction chromatography medium, It was found that the compound binds to the amino acid (Pharmacia). Approximately 50m M phosphorus A phosphate buffer comprising an acid salt and having a pH of about 7.2 is preferred. The ascites is treated with phenylcef. It is applied to a column of allose. Most CA125 is phenyl sepharose It does not bind to the column, flows through the column, and is collected. The column was then adjusted to approximately 2.5 Wash with phosphate buffer containing M urea. This buffer contains the remaining CA1 25 is eluted. The column was then washed with phosphate buffer containing approximately 6M urea. Purify. TPA is eluted with this buffer and recovered.

Chromatography using phenyl sepharose typically allows binding to occur requires high salt concentrations. However, surprisingly, TPA Binds even without high salt concentrations. Thus, this separation is also due to hydrophobic interactions. Since this is not the case, it is normal for this separation to occur. Separated data The protein is buffer exchanged to remove urea and preferably about 1 g 　/　d　Concentrated to higher protein levels than L. Separation of proteins , the isolated proteins were assayed using a commercially available immunoassay method. Therefore, it is confirmed.

CEA, CA19-9 and TPA are often obtained from the same source.

Therefore, they must be separated from each other. CEA is approximately 2゜o,　oo o Dalton large glycoprotein, found at high levels in the serum of colon cancer patients. be discovered. CEA is a carcinofetal antibody that is expressed during intrauterine life and disappears after birth. It is one of the original sources. Carcinoembryonic antigen may be present during pregnancy due to repair or It reappears in conditions of neoplastic growth. lung cancer, stomach cancer, breast cancer, and pancreatic cancer High levels have been found in patients. CA 19-9 is a tumor tumor Chin antigen. Tumor mucin is approximately 200,000 daltons to 1000 k D. A high molecular weight glycoprotein containing about 25% to 80% carbohydrates. . As a tumor marker, CA 19-9 is recommended for patients with pancreatic cancer and gastrointestinal cancer. and is rising.

One of the sources of CEA, CA19-9 and TPAO was specified as 5W1116 It is a cell line. 5W1116 is a human cell line derived from colorectal cancer.

The cancer cells secrete antigen into the cell growth medium.

Collect and freeze the cell growth medium as it is being produced. Thaw the cell supernatant; Concentrate about 20 times. The concentrated supernatant is then dissolved in a solution such as phosphate buffer at physiological pH. Buffer exchange to buffer. A preferred buffer is 50mM phosphate buffer at pH 7.2. It is a liquid solution.

Although CEA, CA19-9 and TPA are somewhat different, they are all is a glycoprotein and is very difficult to separate by typical chromatography. It is difficult to A precipitation method using perchloric acid treatment to precipitate CEA has been proposed. However, with that method, the yield of fft markers is low.

Thus, a method was developed to purify these 3fffi markers.

The concentrated, buffer exchanged supernatant was applied to a phenyl sepharose column. Ru. As described for the purification of 82M, TPA is free from the presence of high concentrations of salts. to the chromatography media.

The column is then washed with phosphate buffer and the eluate is collected in each fraction. Immunoah These fractions contain mostly CA19-9, as determined by However, certain portions contain CEA.

The CEA/CAl9-9 fraction was affinized using methods known in the art. a that can be separated and further purified by tea chromatography, Ford, C , H, J, et al, , Immunoadsorbent Purific ation of Carcinoembryonic Antigen us Ing A Monoclonal Antibody: A Djrect Comparison with a Conventional Method d, Tumor Biol, Vol. 8:p, 241-250 (1987) In the method disclosed in A column containing a feed medium is prepared. This column can remove CEA. Then, CA19-9 passes through the column. CEA can be removed from the column can. However, there are other sources of CEA that are commercially available. Therefore, using this method There is no need to

Since CEA does not need to be obtained in this way, each Combine the fractions and then load the phenyl sepharose column with any additional CA In order to also remove 19-9, a phosphate buffer containing 2 to 3 M urea (preferably It is preferable to wash with 50mM phosphate (pH 7.2). Collect the eluate . Combine CA19-9 and all parts containing CA19-9 and CEA. let The column is then eluted with the same buffer containing 6M urea. TPA elutes and can be collected.

The CAl9-9/CEA-containing pool is buffer exchanged to remove urea and to remove at least about Concentrate to 1 g/dL. Freeze the concentrate. Long-term freezing of CA19-9 binding results in decreased CEA activity, but CA19-9 activity is preserved. . Therefore, the entire purification method can be simplified.

The length of the freezing time will determine the presence of CEA in the immunoassay. This can be determined by testing the parts. Approximate recovery rate can reach 100% .

Alternatively, both parts containing CEA/CA19-9 can be discarded. Next Then, only the fractions containing CA19-9 are pooled and concentrated.

TPA is also buffer exchanged and concentrated as described above. The recovery rate of TPA is also reached approximately 100%. Cross-contamination is measured using immunoassay methods.

CEA can be obtained using monoclonal antibody techniques or from other commercially available sources. can be done. CEA was tested using an immunoassay method prior to use in controls. should be tested for cross-contamination. If contamination is detected, the CE A using one of the methods known in the art, preferably the affinity method described above. It must be purified by

NSE is obtained from fresh or freshly frozen human brain. Purified NSE can be obtained commercially. A preferred purification method involves preparing brain homogenates. , is accomplished by centrifuging the homogenate and collecting the supernatant. next , pellet the supernatant using 40% sodium sulfate. Pellets, 10m resuspended in Tris phosphate buffer of M, dialyzed against the buffer, and then concentrated. do. The concentrate was chromatographed on DE-52 from 0.15M to 0. , 35M sodium chloride gradient. The peak containing MSE Dialyzed, lyophilized and fractionated on Sephadex GI50 etc. Po Rebuffer (Polybuffer) substitution chromatography used to concentrate. The NSE was then eluted and the G-150 ( Finally, fractionation is carried out by Superfine).

Finally, AFP must be purified. AFP, like CEA, is a carcinoembryonic antigen. That's it. AFP is a glycoprotein expressed in the fetal liver and gastrointestinal tract. be. In adults, elevated levels of this antigen in serum are associated with malignant liver cancer. and, in some cases, ovarian cancer and pectinoma. of this antigen The best source is human serum collected at birth. This serum has a high Tumors containing and contaminating Bell's AFP (approximately 60.0 OOn g/mL) Contains only one marker (prolactin).

There are methods for purifying AFP that have been described in the art. For example, Chudy D, and Zizkosky V,, A simple and ra pid method for the 1solation of human alpha-fetoprotein from human cord erum, Neoplasma 34(4), pp, 491-496 (1 987) describes one such procedure. For the purposes of this invention: A preferred method of isolation of AFP from prolactin is ion exchange chromatography. This is accomplished using

Apply the serum to the cation exchange resin using 20mM Tris buffer at pH 8.5. use At this pH and buffer strength, AFP will bind to the column, but to a large extent A portion of prolactin does not bind to the column. In this way, serum is added to the column. The prolactin is then washed out through the column. Prolactin cannot be recovered. I can do it. The AFP is then added to about 0.2 to 0.3 M sodium chloride containing buffer. Thorium can be used to elute from the column.

The isolated AFP is then concentrated to approximately 1 mg/mL. By this method The recovery of AFP can be approximately 100%. AFP purified in this way is , may contain large amounts of albumin. However, this contaminant is not a problem. stomach. This is because the serum or plasma-based material contains albumin. be. Purified AFP can be tested using immunoassay methods.

ACTH, aldosterone, hCG, β-hCG, CA15-3, CA349 , calcitonin, ferritin, gastrin, PAP, PSA, prolactin and Other tumor markers such as LD-1 are commercially available from several sources. child These other markers can be obtained purified or prepared using methods well known in the art. It can be purified by procedures. Immunoassay for each tumor marker can be used to assess cross-contamination. Amount of LD-1 present in LDH By measuring LD-1 is added as a component of LDH.

A control stock solution is used to spike plasma or serum into the plasma or serum. treatment by first assaying all available specific tumor markers. be treated. Table ■ shows the specific tumor mass of each of the three levels of controls prepared. indicates the target value for the marker. The calculation calculates the concentration of each marker in serum or plasma. Subtract from the average target value in Table ■, then add the appropriate amount of each marker to those three levels. to each of the controls.

Tumor markers are added to serum or plasma depending on the stability of each marker. serum or B2MSA as long as the temperature of the plasma is controlled within the range of about 2 to 10'C. FP, prolactin, hCG, β-hCTG, CA15-3, CA19-9 , CA349, CA125, CEA, ferritin, TPA, and LD-1 ( (added as LDH) within a few days of lyophilization. It can be prepared.

If all materials are maintained in the range of approximately 2 to 10"C, ACTH1 gas Thorin, γ-enolase, and calcitonin (a marker with short-term stability in liquid) ) may be added approximately 6 hours prior to lyophilization. Preferably, these markers The car is added just before freeze-drying, and the addition and adjustment are carried out at low temperatures, i.e. It will be held at 10oC.

Each of these three levels of liquid control is lyophilized using standard methods. . The bottles containing the lyophilized controls were sealed under vacuum and then stored at approximately 4°C. Stored. The control is reconstituted with water or other suitable liquid such as a buffer. Ru.

For ACTH, to quantify the loss of ACTH activity during the freeze-drying process. lyophilization studies are required. In order to measure the activity loss due to this process, A non-assay method is used. The results show that retention of specific post-lyophilization levels can be used to determine the pre-lyophilization level of ACTH required for.

The stability of all markers in the lyophilized control was 4.5% at the harsh temperature of 37°C. It was determined that the See Table ■. This is when stored at 2 to 8”C. This corresponds to approximately three years. All markers except ACTH, gastrin and calcitonin Stability after reconstitution of at least two weeks was found for the car. See table ■ . A CT H, gastrin and calcitonin should be reconstituted with liquid immediately. must be used. The stability after reconstitution was determined by NSE, gastrin and calcito. For all analytes other than nin, each aliquot of material after reconstitution was heated to -20°C. It has also been found that the period can be extended for 30 days by freezing. See table About Teru. The stability of gastrin and calcitonin was determined by keeping the amounts of each portion of the material constant after reconstitution. It can be extended for 7 days by freezing at 20'C. See table and. The stability of NSE was determined by freezing each aliquot of the material after reconstitution to -20°C. Therefore, it is possible to extend the time by 24 hours. See Table ■.

The following examples are given to illustrate the invention.

Example l Purification of 82M Urine concentrate was prepared by collecting urine from patients with kidney disease. pool political factions and 0.02% sodium azide was added as a preservative. Removes particles and microorganisms To remove the filtrate, it was filtered through a <0.3 μm membrane. Next, the political faction is pH Diafilter against 7 volumes of 8,0 (50 mM) Lys buffer and the original 100 Concentrated twice. For example, +00 L was concentrated to IL. The concentrated urine is buffered as above. The total protein concentration was adjusted to about 9.0 g/dL using the solution.

Approximately 50 mL of the urea concentrate was applied to an Ultragel ACA 54 column.

Sample size is dependent on column size and equals 2.5% of the total volume of media. Yes. For effective separation of proteins, the column length should be approximately 100 cm. Must be. Fractions containing 82M were combined, pooled and approximately 1 g/d Li: Concentrated.

[H made is also achieved by 5uperdex 75 (Pharmacia) It was done. However, for this application, Llltragel ACA Purification by 54 is superior.

Purification of CA 19-9 and TPA of the supernatant from cell line 5W1116 (supernatant available from Whitaker). Approximately 50 mL of sample was concentrated to half the original volume. 50m with pH approx. 7.2 Samples were buffer exchanged three times with approximately 5 (1 mL) of potassium phosphate at M. . The final volume of the sample was approximately 35 mL. A sample of approximately 20ml, It was applied to a phenyl sepharose column. The column was washed with the buffer and both parts were separated. collected. These fractions were tested for CEA and CA19-9 activity by immunoassay. It was evaluated using the assay. CEA-containing fractions were pooled and concentrated, and CA1 9-9 containing fractions were pooled and concentrated. Approximately 50mM phosphate with gradual increase in urea amount A pH 7.2 buffer containing . TPA is about 6M urea Therefore, it was eluted. The TPA-containing fractions were pooled, concentrated, and then the 6M urine Diafiltration was performed to remove the elements.

To remove any CEA activity contaminating CA 19-9, The A19-9 fraction was subjected to long-term storage.

Example 2 Preparation of controls Defatted serum from the man was filtered through the following filter rows. That is, 1.2μ m pre-filter - 10.8 μm filter, 0.45 μm filter and 0.8 μm filter. It is a 22 μm filter. Filtered serum was refrigerated. Rohm and Ha Broclin 300 from as was added to a concentration of IrnL per serum IL. the blood Divide the liquid into two pools of 1.92L/pool and name them pool 1 and pool 2. I got it.

This serum was combined with ACTH, aldosterone, 82M, hCG, β-h'CG5CA. 15-3, CA 19-9, CAl25, CA349, calcitonin, CE A, ferritin, gastrin, NSE, PAP, PSA, prolactin, TPA , and LD-1 using immunoassay techniques.

Each of these markers was obtained freshly purified as described herein, or Commercially obtained. The amount of each marker was measured. Given in the level I range of table ■ The amount of each marker required to reach the value given was added to Boolean l. Table■ Level ■ Boolean amount of each marker to reach the value given in the range Added to 2. Assay the amount of marker and make any adjustments by adding additional amounts of marker. This was done by addition.

Chill each 3 mL aliquot from Boule 1 (Level I) in the freezer for approximately 1 hour. Fill cooled vials and add each 3 mL aliquot from Boule 2 (Level 3) was filled into vials that had been chilled in the freezer for approximately 1 hour. Freeze the vial Allow to dry, seal under vacuum, and store at approximately 4°C.

Example 3 Assay of Serum Based on Tumor Marker-Control Contains the Control Prepared in Example 2 The vial was reconstituted with 3 mL of distilled water and mixed by gentle inversion. . The markers contained in the level 1 and level 3 controls were It was assayed using the assay method. The results are given in Table IX.

Table 1 Classification of tumor markers 1. Carcinoembryonic antigen AFP, CEA Produced during fetal development and in adults is at a low level. Tumors make use of these proteins. causes the re-emergence of texture.

2. Tumor-associated antigens CA19-9, CA349, CA15-3, whip secreted by tumor cells (carbohydrate-rich glycoprotein). high molecular weight >200 kda, 25-85% carbohydrates 3, hormones hCG, ACTH, calcitonin, protein loractin, aldosterone, gust Rin 4 Serum protein β2-microglobulin, ferritin 5, enzyme PAP, NSE, LD 1 Table 11 Tumor marker comparison clinical markers Tumor marker site Adrenocorticotropic hormone lung (ACTH) α-fetoprotein (AFP) Kanamemaru, liver aldosterone, kidney β2-microglobulin bone marrow β Human Chorionic Gonadotropin Gynecology, Prostate CA 15-3/CA 549 Breast CA 19-9 Pancreas, colorectum, stomach CAl25 Ovary Carcinoembryonic antigen (CEA) colorectal, breast, lung, stomach, pancreas Table 11 (continued) Tumor marker comparison clinical markers Tumor marker site ferritin liver γ enolase lung, brain gastrin pancreas Human chorionic gonadotropin (hCG) Lactate dehydrogenase brain Isoenzyme (LD-1) Prolactin pituitary gland Prostatic acid phosphatase prostate (PAP) Prostate-specific antigen prostate (PSA) Tissue polypeptide antigen Bladder, prostate, gynecology, lung (TPA) Table I11 Tumor marker comparison fresh water CA125 Ri ■ nest cancer ll! 7k, -7 cancer on 0 匈7 Table vI AFP 10.2 10.6 293 294 Aldosterone 79 81 8 04 85082M 1. +1.0 4.2 4.4CA19-9 35 3 3 249 241 Ferritin 31 28 693 671 PAP 2.1 2.0 16.9 16.4 Prolactin 6.8 6.3 +71 170P S A 3.0 2.6 37 32T P A 48 46 953 95 7CA 549 B, 9 8.9 27 30CΔ15-3 41 43 + 65 158βh CG 2.7 2.6 475 446hCG, 2.7 2.7 422 414CAI25 +8 20 356 360CEA 2. 9 2.6, 57 56LDI 47.7 49.8 51 51 Table ■1 Aldosterone 76 78 840 834 AFP 11 11 300 2 97 P2M O,980,974,74,8ACTH1517408418 Ferritin 31 28 726 686P A P 2.2 2. I 20 20P S A 2.6 2.6 33 33 Prolactin 4.2 4.4 125 133T P A 56 54 779 854CA549 9.1 9.9 36 35CA15-3 23 23 100 102βh CG 2.6 2.6 426 438CA125 25 25 448 474CE A 2.6 2.6 60 6I CA19-9 55 55 276 277L D H25224775676 4 LD-148% 48% 51% 51% table ■ Gastrin 101 98 322 313 Calcitonin 123 110 3 42 353 NSE 10 10 54 54 Table 1× Equipment/method comparison Analysis target Method Unit Level 1 Level 3 Manufactured by ACTH Diagnostics Product pg/mL 33 461 [ncstar RIA pg/mL 14 4 66 Nichols Allegro RIA pmol/L 14 69C1i nical As5ays pg/mI, 20 466 aldosterone Diagnostics product pg/mL 79 875 β2-microglobe Rin CA125° Centocor U/mL 22 377CA349)IY britech Tandem-RU/mL 10 30 Table IX (continued) Equipment/method comparison Gastrin C11nical As5ays pg/mL 172 46°D iagnostics products pg/mL 56 347 Table IX (continued) Equipment/method comparison TP A BYk-5angtec ng/mL 54 978 Calcitonin Diagnostics product pg/mL 61 1731ncstar, I [RIA pg/mL 129 367*CA15-3, CA 19-9, C A125 is Centocor DJagnostr, a division of Centocor. It is a registered trademark of cs.

Claims (2)

[Claims] 1. Controls for the measurement of tumor markers, comprising (a) lipid-reduced human A base comprising serum or plasma, wherein the serum or plasma essentially contains antibodies against PAP. A base that does not include (b) multiple tumor markers; A control containing a mixture of 2. The control of claim 1, which is lyophilized.