sepharose and Colonic-Neoplasms

High-fat diet (HFD)-induced alteration in the gut microbial composition, known as dysbiosis, is increasingly recognized as a major risk factor for various diseases, including colon cancer. This report describes a comprehensive investigation of the effect of agaro-oligosaccharides (AGO) on HFD-induced gut dysbiosis, including alterations in short-chain fatty acid contents and bile acid metabolism in mice. C57BL/6N mice were fed a control diet or HFD, with or without AGO. Terminal restriction fragment-length polymorphism (T-RFLP) analysis produced their fecal microbiota profiles. Profiles of cecal organic acids and serum bile acids were determined, respectively, using HPLC and liquid chromatography-tandem mass spectrometry systems. T-RFLP analyses showed that an HFD changed the gut microbiota significantly. Changes in the microbiota composition induced by an HFD were characterized by a decrease in the order Lactobacillales and by an increase in the Clostridium subcluster XIVa. These changes of the microbiota community generated by HFD treatment were suppressed by AGO supplementation. As supported by the data of the proportion of Lactobacillales order, the concentration of lactic acid increased in the HFD + AGO group. Data from the serum bile acid profile showed that the level of deoxycholic acid, a carcinogenic secondary bile acid produced by gut bacteria, was increased in HFD-receiving mice. The upregulation tended to be suppressed by AGO supplementation. Finally, results show that AGO supplementation suppressed the azoxymethane-induced generation of aberrant crypt foci in the colon derived from HFD-treated mice. Our results suggest that oral intake of AGO prevents HFD-induced gut dysbiosis, thereby inhibiting colon carcinogenesis.

Topics: Animals; Bile Acids and Salts; Clostridium; Colonic Neoplasms; Diet, High-Fat; Dietary Fiber; Dysbiosis; Endotoxins; Fatty Acids; Feces; Lactobacillales; Male; Mice; Mice, Inbred C57BL; Microbiota; Obesity; Oligosaccharides; Sepharose

Topics: Animals; Colonic Neoplasms; Diet, High-Fat; Dysbiosis; Male; Oligosaccharides; Sepharose

Innovative diagnostic methods are the need of the hour that could complement conventional histopathology for cancer diagnosis. In this perspective, we propose a new concept based on spectral histopathology, using IR spectral micro-imaging, directly applied to paraffinized colon tissue array stabilized in an agarose matrix without any chemical pre-treatment. In order to correct spectral interferences from paraffin and agarose, a mathematical procedure is implemented. The corrected spectral images are then processed by a multivariate clustering method to automatically recover, on the basis of their intrinsic molecular composition, the main histological classes of the normal and the tumoral colon tissue. The spectral signatures from different histological classes of the colonic tissues are analyzed using statistical methods (Kruskal-Wallis test and principal component analysis) to identify the most discriminant IR features. These features allow characterizing some of the biomolecular alterations associated with malignancy. Thus, via a single analysis, in a label-free and nondestructive manner, main changes associated with nucleotide, carbohydrates, and collagen features can be identified simultaneously between the compared normal and the cancerous tissues. The present study demonstrates the potential of IR spectral imaging as a complementary modern tool, to conventional histopathology, for an objective cancer diagnosis directly from paraffin-embedded tissue arrays.

Topics: Adenocarcinoma; Algorithms; Carbohydrates; Collagen; Colonic Neoplasms; Diagnostic Imaging; Humans; Nucleotides; Optical Phenomena; Paraffin Embedding; Principal Component Analysis; Sepharose; Spectrophotometry, Infrared; Spectroscopy, Fourier Transform Infrared; Tissue Array Analysis

For over a decade our laboratory has developed and used a novel histochemical assay using derivatized agarose beads to examine the surface properties of various cell types. Most recently, we have used this assay to examine lectin binding ligands on two human cell types, CCL-220, a colon cancer cell line, and CRL-1459, a non-cancer colon cell line. We found that CCL-220 cells bound specific lectins better than CRL-1459, and this information was used to test for possible differential toxicity of these lectins in culture, as a possible approach in the design of more specific anti-cancer drugs. Although we have examined the validity of the bead-binding assay in sea urchin cell systems, we have not previously validated this technique for mammalian cells. Here the binding results of the bead assay are compared with conventional fluorescence assays, using lectins from three species (Triticum vulgaris, Phaseolus vulgaris, and Lens culinaris) on the two colon cell lines. These lectins were chosen because they seemed to interact with the two cell lines differently. Binding results obtained using both assays were compared for frozen, thawed and fixed; cultured and fixed; and live cells. Both qualitative and quantitative fluorescence results generally correlated with those using the bead assay. Similar results were also obtained with all of the three different cell preparation protocols. The fluorescence assay was able to detect lower lectin binding ligand levels than the bead assay, while the bead assay, because it can so rapidly detect cells with large numbers of lectin binding ligands, is ideal for initial screening studies that seek to identify cells that are rich in surface binders for specific molecules. The direct use of frozen, thawed and fixed cells allows rapid mass screening for surface molecules, without the requirement for costly and time consuming cell culture.

Topics: Cell Line; Cell Line, Tumor; Colon; Colonic Neoplasms; Fluorometry; Histocytochemistry; Humans; Lectins; Ligands; Membrane Proteins; Microspheres; Phytohemagglutinins; Plant Lectins; Protein Binding; Reproducibility of Results; Sepharose; Wheat Germ Agglutinins

Matrilysin produced by human colon cancer cells may be involved in the progression and metastasis of cancer. In the present study, we investigated the association of matrilysin with angiogenesis. One microgram of recombinant matrilysin is confirmed to have increased [3H]-thymidine uptake in human umbilical vein endothelial cells. Then we used micro encapsulation and a mouse hemoglobin enzyme-linked immunosorbent assay system for in vivo quantitation of angiogenesis with BALB/c nu/nu athymic mice. Hundred micrograms of recombinant matrilysin induced angiogenesis to the same degree as 10 microg of basic fibroblast growth factor (bFGF). Angiogenesis was observed at the site implanted with human colon cancer WiDr cells in agarose micro beads. This was inhibited by subcutaneous injection of matrilysin-specific antisense oligonucleotide significantly by 53%. In conclusion, matrilysin may be associated with angiogenesis of human colon cancer through the direct proliferative action on endothelial cells.

Topics: Animals; Cell Division; Cells, Cultured; Colonic Neoplasms; Colorectal Neoplasms; Culture Media, Serum-Free; DNA; Endothelium, Vascular; Enzyme-Linked Immunosorbent Assay; Fibroblast Growth Factor 2; Humans; Immunoblotting; Matrix Metalloproteinase 7; Mice; Mice, Inbred BALB C; Neoplasm Metastasis; Neoplasm Transplantation; Neovascularization, Pathologic; Oligonucleotides, Antisense; Sepharose; Umbilical Veins

Human colon cancer cell lines express epidermal growth factor (EGF) mRNA, secrete EGF and may respond to it via the cell-surface EGF receptor (EGFR). Expression of these molecules in human colon and colon tumor, however, is not clear. Reverse transcription-polymerase chain reaction (RT-PCR) analyses of RNA prepared from paired normal human colon and colon tumor samples from 12 individuals followed by Southern blotting analyses of the RT-PCR products revealed a major fragment of 527 bp and a minor fragment of 404 bp that hybridized to a human EGF cDNA probe under stringent conditions. Identical results were obtained from 8 human colon cancer cell lines. Cloning and sequencing of PCR products confirmed that both fragments were from the human EGF gene; the 527-bp fragment corresponded exactly to nucleotides 2,891 to 3,417 of the human EGF mRNA reported by others. A deletion of 123 nucleotides (nucleotides 3,172 to 3,294) was found in the 404-bp fragment. Immunohistochemical studies using cyostat sections of human colon specimens showed that EGF was expressed in the human colon and that expression was restricted to the epithelial colonic crypt cells and epithelium-derived cancer cells. Since EGF and EGF-related molecules are potent mitogens that mediated their effect through the EGFR, we also determined the efficacy of anti-sense EGFR RNA in circumventing the EGFR-related pathway of proliferation. Expression of anti-sense EGFR RNA, by transfection with an inducible anti-sense EGFR expression vector, down-regulated cell-surface EGFR expression and proliferation of these cells and their ability to grow in soft agar. Anti-sense EGFR RNA was found to be an anti-proliferative agent in both relatively non-aggressive and highly aggressive human colon cancer cells.

Topics: Adenocarcinoma; Base Sequence; Blotting, Southern; Cell Division; Cloning, Molecular; Colon; Colonic Neoplasms; Down-Regulation; Epidermal Growth Factor; Epithelium; ErbB Receptors; Humans; Molecular Sequence Data; Polymerase Chain Reaction; RNA, Antisense; RNA, Messenger; Sepharose; Transcription, Genetic; Transfection; Tumor Cells, Cultured

A gel electrophoretic method coupled with agar diffusion has been devised for detecting tumor antigens in human colon tissue. Separation of the antigens is achieved on duplicate electrophoretic gels. One gel is used for the location of the antigens by protein staining and the other gel is used for assaying of the antigenicity by agar diffusion against homologous antiserum. Analysis of perchloric acid extracts of colon tumors by this coupled method revealed the presence of carcinoembryonic antigen and two additional glycoprotein antigens. Analysis of KCl-HCl tumor extracts revealed two new tumor antigens.

Topics: Adenocarcinoma; Antigens, Neoplasm; Carcinoembryonic Antigen; Colonic Neoplasms; Electrophoresis, Polyacrylamide Gel; Humans; Immunodiffusion; Perchlorates; Sepharose

Human colon cancer (Moser) cells produce and secrete epidermal growth factor (EGF) and respond to EGF via an autocrine/paracrine mode through the cell surface EGF receptor (EGFR). In this report we show that EGF promotes the malignant behavior of the Moser cells in vitro in terms of growth in soft agarose and invasion of Matrigel-coated porous membranes. Expressing antisense EGFR RNA in the Moser cells (through transfection with an inducible antisense EGFR expression vector) downmodulated the expression of cell surface EGFR and EGFR mRNA with a concurrent inhibition of growth in soft agarose and invasion of Matrigel-coated membranes. In addition, the ability of exogenously applied EGF in promoting the malignant behavior of these cells was circumvented. We conclude that antisense EGFR RNA was a potent agent in circumventing the in vitro malignant properties of the Moser cells.

Topics: Cell Division; Collagen; Colonic Neoplasms; Down-Regulation; Drug Combinations; Epidermal Growth Factor; ErbB Receptors; Gene Expression; Gene Expression Regulation, Neoplastic; Humans; Laminin; Neoplasm Invasiveness; Proteoglycans; RNA, Antisense; RNA, Messenger; Sepharose; Transfection; Tumor Cells, Cultured

A cleaning resin has been developed for the non-chromatographic purification of 99mTc-labeled monoclonal antibodies (MAbs). The resin used is a modified form of thiopropylsepharose 6B resin, in which its sulfhydryl groups have been tinylated with stannous chloride. The method requires only simple stirring of the radiolabeling reaction mixture with this tinylated resin and subsequent separation of it from the resin by filtration to obtain a 99mTc-labeled MAb of radiopharmaceutical purity. The method provides an alternative to chromatographic purification of the radiolabeled MAb (i.e. gel filtration or anion exchange chromatography) which has been used in other 99mTc-MAb preparations. For comparison studies, we labeled the B72.3 MAb with NeoRx's diamide dimercaptide chelate radiolabeling kit, split the reaction mixture into two equal portions and then purified one portion with anion exchange chromatography (NeoRx's chosen method) while the other portion was purified with our cleaning resin. Comparison of HPLC chromatograms, percent 99mTc-bound to MAb, biodistribution and scintigraphic results show that our cleaning resin methodology provides a 99mTc-labeled MAb of essentially equal purity and utility as does the established, chromatographic one.

Topics: Animals; Antibodies, Monoclonal; Colonic Neoplasms; Humans; Mice; Mice, Nude; Molecular Structure; Radionuclide Imaging; Radiopharmaceuticals; Resins, Plant; Sepharose; Technetium; Tissue Distribution; Transplantation, Heterologous

Direct tissue isoelectric focusing was used as a procedure to analyze differences in soluble tissue protein profiles of resected intestinal segments and endoscopic biopsies from patients with ulcerative colitis, Crohn's disease, and colonic cancer. Extraction of tissue proteins was accomplished by electrophoresis of mucosal cryostat sections on agarose gels across a broad pH gradient. The inflamed colonic mucosa from Crohn's disease patients showed similar isoelectric focusing protein patterns. Small bowel mucosa from a patient with both colonic diverticular disease and Crohn's disease showed protein patterns identical with that of the mucosa from a patient with only Crohn's disease. The inflamed mucosae from ulcerative colitis patients revealed identical protein patterns but were distinct from those of non-inflamed ulcerative colitis mucosa and from the inflamed mucosae from Crohn's disease patients. Non-inflamed small bowel mucosae from cancer, ulcerative colitis, and Crohn's disease patients showed distinct protein patterns which were absent in the non-inflamed large bowel mucosae. The inflamed resected ileum of a Crohn's disease patient exhibited protein patterns similar to those of the biopsy of an inflamed mid-transverse large bowel. Mucosal biopsies from inflamed sigmoid colon of a Crohn's disease patient showed different protein patterns than those in biopsies from the inflamed mid-transverse colon. Thus, distinctive isoelectric focusing protein patterns may be useful in differentiating Crohn's colitis and ulcerative colitis when granulomata are absent, and in resolving indeterminant colitis to one of these classic inflammatory bowel diseases.

Topics: Biopsy; Colitis, Ulcerative; Colon; Colonic Neoplasms; Crohn Disease; Epitopes; Humans; Inflammatory Bowel Diseases; Intestinal Mucosa; Isoelectric Focusing; Neoplasm Proteins; Proteins; Sepharose

Insulin-like growth factor I (IGF-I) stimulates the proliferation of highly metastatic NL-17 cells to a greater extent than poorly metastatic NL-44 cells, both of which are derived from mouse colon carcinoma 26. The NL-17 cells have been compared with NL-44 cells for the signal transduction pathway of IGF-I. IGF-I receptors of both cell types were identified by affinity labeling, and there was no significant difference between the two cell types in the amount or the autophosphorylation activity of the IGF-I receptors. However, when IGF-I-dependent tyrosine phosphorylation of cellular components was examined, remarkable tyrosine phosphorylation of proteins with molecular weights of 150,000 (pp150) and 160,000 (pp160) was found in NL-17 cells. In contrast, this phosphorylation stayed at significantly lower levels in NL-44 cells than in NL-17 cells. The phosphorylation of pp150 and pp160 was induced within 10 s after the addition of IGF-I and reached its maximal level by 30 s. After the removal of IGF-I, the phosphorylation of pp150 and pp160 was reduced to the basal level within 30 min. This phosphorylation was not induced by platelet-derived or epidermal growth factor. The pp150 and pp160 were not absorbed by wheat germ agglutinin-agarose. They were found in the soluble fraction of cytoplasm but not in the membrane or the cytoskeleton. The pp150 and pp160 might be endogenous substrates of IGF-I receptor kinase. These results suggest that tyrosine phosphorylation of pp150 and pp160 mediates the higher proliferative response of NL-17 cells to IGF-I.

Topics: Animals; Antibodies, Monoclonal; Cell Line; Colonic Neoplasms; Epidermal Growth Factor; Insulin; Insulin-Like Growth Factor I; Kinetics; Mice; Molecular Weight; Neoplasm Metastasis; Neoplasm Proteins; Phosphoproteins; Phosphorylation; Phosphotyrosine; Platelet-Derived Growth Factor; Receptors, Cell Surface; Receptors, Somatomedin; Sepharose; Tyrosine

The purpose of this study was to determine whether the degree of anchorage-independent growth of human tumor cells in increasing concentrations of agarose correlated with the capacity of the cells to produce experimental metastases in nude mice. Human melanoma, breast carcinoma, and colon carcinoma cells from parental lines and variants selected in vivo for metastasis and in vitro cloned lines were plated into medium containing 0.3%, 0.6%, 0.9%, or 1.2% of agarose. These cells were also injected into nude mice: intravenously for melanoma, into the mammary fat pad for breast carcinoma, and into the spleen for colon carcinoma. Production of tumor cell colonies in dense agarose (greater than 0.6%) correlated with production of experimental metastases in the lung (melanoma, breast carcinoma) or liver (colon carcinoma). We conclude that the degree of anchorage-independent growth of tumor cells can predict their biological behavior and metastatic potential in vivo. Thus, this technique may be useful for the isolation of metastatic cells from heterogeneous human neoplasms.

Topics: Animals; Breast Neoplasms; Clone Cells; Colonic Neoplasms; Culture Media; Humans; Lung Neoplasms; Melanoma; Mice; Mice, Nude; Neoplasm Metastasis; Neoplasm Transplantation; Sepharose; Tumor Cells, Cultured

The human colon, intratumoral subpopulations HCT 116 and HCT 116a were established in chemically defined medium supplemented with transferrin, insulin, epidermal growth factor (EGF), triiodothyronine, hydrocortisone, and sodium selenite. The responsiveness of the adapted cell lines to these growth factors was compared in anchorage-dependent and -independent assays. HCT 116 cells maintained in serum-free conditions were further adapted to growth factor deprivation, and the effects of these polypeptides were determined in anchorage-independent assays. In monolayer, HCT 116 cells adapted to grow in serum-free medium responded to transferrin but not to EGF or insulin. Similarly adapted HCT 116a cells were, however, insensitive to transferrin addition but manifested a 300 and 500% increase in growth rates with EGF and insulin, respectively. Optimal growth of HCT 116 cells was seen in the presence of insulin and transferrin, while maximum proliferation of HCT 116a cells depended on combined insulin, transferrin, and EGF. In soft agarose, both HCT 116 and HCT 116a subpopulations showed a stringent requirement for transferrin. No combination of growth factors without transferrin supported colony formation. These data suggest that (a) these colon tumor subpopulations may be subject to separate growth controls, and (b) there may be an important role for transferrin in anchorage-independent growth and possibly in the maintenance of malignant characteristics.

Topics: Cell Division; Colonic Neoplasms; Culture Media; Epidermal Growth Factor; Humans; Insulin; Sepharose; Suspensions; Transferrin; Tumor Cells, Cultured

The feasibility of extracorporeal adsorption of 1.5-3 L plasma on protein A-Sepharose was investigated in six patients with advanced cancer. Anticoagulation with heparin was associated with respiratory distress syndrome in two patients, most likely caused by complement activation as indicated by a transient leukopenia during plasma reinfusion and appearance of C3 degradation products in the extracorporeal circulation. Addition of citrate abolished the respiratory symptoms, C3 degradation, and leukopenia, and no adverse reactions were observed. No objective tumor regression was observed in any of the patients. Three patients progressed during therapy. In one of these, multifocal central tumor necrosis was observed as a possible, although unproven, therapeutic effect. Increased natural killer and/or killer cell activities were recorded in three patients and increased complement-dependent serum cytotoxicity in one patient. The level of circulating immune complexes decreased significantly (18-28%) in three patients studied. It is concluded that extracorporeal plasma adsorption on protein A-Sepharose is feasible when citrate is added to the extracorporeal system, but its therapeutic efficacy is uncertain.

Topics: Adsorption; Adult; Aged; Colonic Neoplasms; Complement Activation; Complement C3; Female; Humans; Immunoelectrophoresis, Two-Dimensional; Immunosorbent Techniques; Kidney Neoplasms; Killer Cells, Natural; Male; Melanoma; Middle Aged; Renal Dialysis; Sepharose; Staphylococcal Protein A

Purification of radiolabeled carcinoembryonic antigen (CEA) preparations by affinity chromatography with anti-AG bound to Sepharose was attempted, since an immunological similarity between AG (alpha 1-acid glycoprotein) and a portion of CEA had been noted. When 125I-CEA was purified in this manner, the fraction which did not bind to the column showed decreased reactivity with either anti-AG or anti-CEA. The retained fraction showed enhanced reactivity with both anti-AG and anti-CEA. The yield of purified CEA increased when the CEA preparation was allowed to react with the anti-AG column overnight. Purification of CEA from tumor tissue was performed by affinity chromatography. A perchloric acid (PCA) extract from cancer tissue was mixed with antiserum against CEA to give an immune complex, and a CEA-reactive fraction obtained by PCA extraction. The CEA-reactive fraction was eluted from a Sephadex G-200 column, and final purification was by anti-AG chromatography. When purified CEA was applied to a Sephadex G-200 column with carrier protein after labeling with 125I, the eluted radioactivity was found only in the 180,000 dalton fraction. Almost all the radioactivity was precipitated from the labeled protein by either anti-AG or anti-CEA. Purification of CEA is possible by affinity chromatography with anti-AG bound to Sepharose.

Topics: Animals; Carcinoembryonic Antigen; Chromatography, Affinity; Chromatography, Gel; Colonic Neoplasms; Humans; Immune Sera; Orosomucoid; Rabbits; Sepharose

A soft agarose clonogenic assay is presented which has been optimized for the growth of human gastrointestinal adenocarcinomas. Samples from 15 gastric and colonic solid tumors and from 2 noncancerous stomachs (control cultures) were disaggregated by treatment with collagenase at 37 degrees overnight. Colonies appeared 10 to 15 days after plating, with a cloning efficiency between 0 and 0.82%, which was markedly improved by a fibroblastic feeder layer. The results suggest a correlation between cloning efficiency and the degree of differentiation of the initial tumor. Histochemistry, electron microscopy, and a carcinoembryonic antigen immunofluorescence assay showed that the colonies consisted of cells with the same characteristics as those of the original tumor.. This colony formation assay appears to be potentially useful for assessing the stem cell pool of gastrointestinal tumors. It will be valuable for studying their response to chemotherapeutic agents in vitro. This clonogenic assay may also permit the establishment of cancer cell lines.

Topics: Adenocarcinoma; Carcinoembryonic Antigen; Cell Differentiation; Cell Division; Cell Line; Cells, Cultured; Colonic Neoplasms; Colony-Forming Units Assay; Gastrointestinal Neoplasms; Humans; Sepharose; Stomach Neoplasms

We are isoelectric focusing small amounts of tissues placed directly on electroendosmosis-free agarose gels instead of using the saline extracts of tissue homogenates. More numerous proteins are extracted during isoelectric focusing of the solid tissues than present in the same tissues.

Topics: Adenocarcinoma; Colonic Neoplasms; Histological Techniques; Humans; Isoelectric Focusing; Neoplasm Proteins; Sepharose; Time Factors

We describe here the use of a new agarose for isoelectric focusing of body fluids and tissue proteins. Macromolecular proteins even when significantly greater in size than 1 X 10(6) mol.wt. were easily and rapidly focused.

Topics: Adenocarcinoma; Ascitic Fluid; Blood Proteins; Colonic Neoplasms; Electrophoresis, Polyacrylamide Gel; Humans; Isoelectric Focusing; Neoplasm Proteins; Polysaccharides; Sepharose

Leukocyte migration tests under agarose (Clausen technique) were performed in 28 patients tentatively diagnosed as having any malignancy with the use of a 3 M KCl-extract panel prepared from bronchogenic, gastric, colonic, renal, and mammary carcinoma, corresponding normal tissues, carcinoembryonic antigen (CEA), and human encephalitogenic protein (HEP). 17 out of 22 proven carcinoma patients showed sensitization by reaction with optimal concentrated KCl-extract of cancer from the same organ type as their own tumor. In some cases positive reactions could be observed also with normal tissue antigen (NTA) of tumor organ type (7/22) or with an additional carcinoma extract of organ type differing from patients own primary tumor (8/22). Gastrointestinal carcinomas, especially, showed sensitization to CEA (7/12) contrary to nongastrointestinal carcinomas (1/10). With HEP no positive reactivity could be found (0/10). With the use of tumor antigen panel (5 antigens) only few positive reactions (MI less than 0.80 or greater than 1.20) could be observed in 6 patients with nonmalignant diseases (1/30 tests) and 8 healthy blood donors (1/40 tests). A widespread individual screening program using tissue antigens for patients suspected of malignancies could give a pattern of reactivities and improve the recognition of cell-mediated sensitization against tumor tissues.

Topics: Aged; Antigens, Neoplasm; Breast Neoplasms; Carcinoembryonic Antigen; Cell Migration Inhibition; Colonic Neoplasms; Epitopes; Female; Humans; Immunity, Cellular; Kidney Neoplasms; Leukocytes; Lung Neoplasms; Male; Middle Aged; Myelin Basic Protein; Neoplasms; Sepharose; Stomach Neoplasms

Topics: Adenocarcinoma; Animals; Binding Sites, Antibody; Carcinoembryonic Antigen; Chromatography, Affinity; Colonic Neoplasms; Goats; Humans; Immunoelectrophoresis; Immunoglobulin G; Iodine Radioisotopes; Liver Neoplasms; Neoplasm Metastasis; Radioimmunoassay; Sepharose

Topics: Adenocarcinoma; Aluminum Hydroxide; Animals; Animals, Newborn; Antibody Formation; Carcinoembryonic Antigen; Colonic Neoplasms; Goats; Horses; Immune Sera; Immune Tolerance; Immunization, Secondary; Immunodiffusion; Immunoelectrophoresis; Liver Neoplasms; Neoplasm Metastasis; Rabbits; Radioimmunoassay; Sepharose