Monoclonal Antibodies For Diagnosis Research Articles

Aptamer as a proper alternative instead of monoclonal antibody in diagnosis and neutralization of menacing biological agents

Abstract: Of the major threats to contemporary mankind, is the use of very dangerous and lethal biological agents as the biological weapons. The first step in confronting with this serious threat after prevention, is the accurate and rapid detection of this agents and neutralization of them. In this article, the role of molecules known as aptamer, has been studied in biological defense against these menacing biological agents. Traditional methods for detection of these agents are based primarily on immuno-affinity assays and the use of antibody molecules. While the modern methods, based on aptamer-affinity assays, are being replaced with traditional methods, due to the abundant advantages of them. The selection and preparation method of specific aptamer with high binding affinity to these biological agents is known as SELEX and the use of magnetic nanoparticles to perform this procedure (Mag-SELEX) is very common. The isolated aptamers with high specificity can also be used in neutralization and inhibition of menacing agents function, in addition to, quick and accurate diagnosis of these agents, utilizing them in nano-biosensors, based on aptamers (as the nano-aptasensors). Keywords: Aptamer, SELEX, nano aptasensor, diagnosis and neutralization, menacing biological agents.

P89 - Comparison between modified acid fast stain and antigen detection assay by monoclonal antibody in diagnosis of Cryptosporidium parvum

P89 - Comparison between modified acid fast stain and antigen detection assay by monoclonal antibody in diagnosis of Cryptosporidium parvum

Combined detection of pepsinogen and gastric cancer monoclonal antibody for diagnosis of precancerous lesions of gastric cancer

Combined detection of pepsinogen and gastric cancer monoclonal antibody for diagnosis of precancerous lesions of gastric cancer

Monoclonal Antibodies for Diagnosis of Enterovirus 71

Enterovirus 71 (EV71), one of the major causative agents of hand, foot, and mouth disease (HFMD), is now recognized as an emerging neurotropic virus in Asia and may cause severe neurologic complications and mortalities. Laboratory diagnosis of EV71 infection must be efficient and accurate, which could be accomplished by various immunoassays. In this study we use a live EV71 isolate, Tainan/4643/98, with genotype C2 as an immunogen to sensitize BALB/c (H-2(d)) mice and then generate the EV71-specific murine monoclonal antibodies. Five hybridoma clones were established and their monoclonal antibodies were characterized. All five clones are applicable in immunofluorescence staining but with different sensitivities-that is, MAbs 22, 24, and 27 were sensitive in IFA detection, and MAbs 22 and 24 were also confirmed in flow cytometry. None of these cross-reacted with coxsackievirus A16 (CVA16) or Echovirus type 6 (ECHO6), but each varied in binding to different EV71 subgenogroups (B1, B4, B5, C2, and C4). Western blot analysis revealed that all of these MAbs reacted with EV71 VP1 capsid proteins, and in addition MAbs 22 and 24 exhibited potent neutralizing activities against EV71 and protected cells from infection. Further, mapping the epitopes for each MAb revealed that only MAb 27 showed positive for the linear epitope DVIESSIGDSVSRAL, which was located at the N-terminus (a.a. 6-20) of EV71 VP1 and highly conserved among all EV71 subgenotypes. Thus, these MAbs may provide valuable tools for the laboratory diagnosis of EV71 infection and for vaccine development.

Value of Golgi protein 73 monoclonal antibody in diagnosis of hepatocellular carcinoma.

To explore the sensitivity and specificity of Golgi protein 73 (GP73) monoclonal antibody in the diagnosis of hepatocellular carcinoma (HCC). Self-prepared GP73 monoclonal antibody was used as the primary antibody for detecting the serum GP73 levels in healthy controls(n=31)and HCC patients (n=59). The baseline level of the healthy controls was determined by semiquantitative analysis. The results were compared with those from GP73 polyclonal antibody and alpha-fetoprotein (AFP). The GP73 level of healthy controls was 1.2 (0.9-1.7) relative unit (RU), which was significantly lower than that of HCC patients [5.7 (2.5-7.8) RU] (P<0.001) with monoclonal antibody. Using polyclonal antibody, the GP73 level of HCC patients was also significantly higher than healthy controls [7.8 (3.0-12.4) RU vs. 1.1 (1.0-2.0) RU, P<0.001]. The sensitivity and specificity of GP73 monoclonal antibody in diagnosis of HCC were 84.7% and 93.5%; on the contrary, those of GP73 polyclonal antibody were 78.0% and 93.5%, respectively. The sensitivity and specificity of AFP (67.8% and 74.2%, respectively) in the HCC patients were markedly lower than those of GP73. Logistic regression analysis showed that the odds ratio (OR) of GP73 monoclonal antibody was 7.18 and that of GP73 polyclonal antibody was 1.51. Our self-prepared monoclonal antibody can effectively detect GP73 serum level in HCC patients, and has higher sensitivity and specificity than AFP. It may be superior to the currently used GP73 polyclonal antibody. The results lay the foundation for the further development of ELISA methods by using this monoclonal antibody.

Monoclonal antibodies in the diagnosis of lymphoproliferative diseases of the orbit and orbital adnexae

To describe the value of monoclonal antibodies in the differential diagnosis of orbital lymphoproliferative disorders. A total of 200 sequential cases of malignant lymphoma diagnosed at St Bartholomew's Hospital as part of the ocular lymphoma service at Moorfields Eye Hospital and the Institute of Ophthalmology have been examined. Cases were acquired between January 1998 and June 2005. Each case had detailed immunophenotypic analysis using a panel of monoclonal antibodies and was classified using the WHO classification of lymphoma. These cases are discussed in relation to earlier studies of orbital lymphoma reported by our group. Lymphomas fell into three main categories. Extranodal marginal zone lymphoma was the largest group with 151 cases, arising within the orbital soft tissue, conjunctiva and lacrimal gland. Cases arising in the conjunctiva and lacrimal gland showed a higher female predominance than those arising within the deeper soft tissue. A small number of cases were associated with organ specific autoimmunity, including thyroid eye disease complicating Graves' disease. Follicular lymphoma and diffuse large B-cell lymphoma formed the next two groups, occurring with equal frequency. Many of the follicular lymphomas had evidence of disseminated disease on completion of staging. A miscellaneous group of T-cell and B-cell lymphomas formed a minority of cases during the study period. Extranodal marginal zone lymphoma is the most frequent type of primary orbital and orbital adnexal lymphoma. Its major differential diagnosis is with orbital lymphoid hyperplasia, chronic dacryoadenitis, and follicular conjunctivitis. Systemic types of lymphoma may present within the orbit or involve the orbit secondarily.

Use of a novel monoclonal antibody in diagnosis of Barrett's esophagus.

A novel monoclonal antibody (MAbDAS-1), that specifically reacts with colonic but not small intestinal epithelium, recognizes specialized columnar epithelium (SCE) in the esophagus. The frequency of its reactivity in biopsy specimens of patients with endoscopically suspected Barrett's Esophagus (BE) is examined. Fifty-two biopsy specimens of the distal esophagus from 38 patients were tested by immunoperoxidase method using MAbDAS-1. Fifty-four samples of cardia-type mucosa biopsied from the stomach were used as controls. Results were compared with histology and Alcian blue/high iron diamine (AB/HID). Of the 52 specimens, 29 had glandular epithelium and the rest had only squamous epithelium. Ten were diagnosed to have SCE by histology. All 10 samples reacted with MAbDAS-1 and with Alcian blue. Of the remaining 19 specimens, five also reacted with MAbDAS-1. None of the squamous epithelium and cardia specimens reacted with MAbDAS-1. MAbDAS-1 may detect intestinal metaplasia of the esophagus of colonic phenotype in the absence of histological evidence of SCE.

4th Scandinavian Symposium on Radiolabeled Monoclonal Antibodies in Diagnosis and Therapy of Cancer

4th Scandinavian Symposium on Radiolabeled Monoclonal Antibodies in Diagnosis and Therapy of Cancer

Summary and Perspectives

The papers of the 4th Scandinavian Symposium on Radiolabeled Monoclonal Antibodies in Diagnosis and Therapy of Cancer are summarised. Particular emphasis is placed on quantitation, dosimetry and radionuclide therapy. Some biological aspects of radionuclide therapy are indicated.

Monoclonal antibodies for diagnosis and therapy of squamous cell carcinoma of the head and neck.

O grupo de carcinomas espinocelulares da cabeca e pescoco em pacientes com alto risco de desenvolver metastases a distância, tem sido bem definidos do ponto de vista prognostico em nossa instituicao. Com intuito de selecionar fatores de risco preditivos no desenvolvimento de metastases a distância, Leemans e col analisaram um grupo de 281 pacientes com carcinoma epidermoide que nao desenvolveram doenca recorrente acima da clavicula (79). Todos os pacientes foram primariamente operados e submetidos a radioterapia complementar diante de 3 ou mais linfonodos histologicos metastaticos na avaliacao da peca operatoria, sendo que neste grupo, a incidencia de metastases a distância a 5 anos foi de 10,7%. 0 numero de metastases em Iinfonodos em n4 de 3 ou mais, foi determinante do desenvolvimento de metastases a distância em 50% dos casos, fato este nao detectado quando esta cifra estava abaixo de 3 linfonodopatias. Em contrapartida, a presenca de ruptura extra-capsular foi um fator dramatico de mudanca de prognostico no que diz respeito a presenca de metastases a distância (3 vezes mais que nos casos onde a ruptura extra-capsular estava ausente). Outro aspecto que mereceu a atencao dos autores, foi aquele relacionado com os pacientes recidivados loco-regionalmente ocorrendo nestes pacientes duas vezes mais metastases sistemicas (80,81). A opcao proposta no sentido de introduzir uma terapia adjuvante para os pacientes com alto risco de desenvolver metastases a distância, e o emprego de anticorpos monoclonais marcados. Recentemente, apresentamos um primeiro estudo experimental em carcinoma epidermoide da cabeca e pescoco tratados por radio-imunoterapia. Foi demonstrado que a IgG E48 de anticorpos monoclonais marcados com 1131 foi capaz de eliminar implantes humanos de carcinoma escamocelular em tumores transplantados para ratos(82). Em 20% dos experimentos desenvolvidos (2 em 10 tumores), foi observada uma regressao em ratos tratados com 400iCi de IgG E48 de anticorpos monoclonais marcados com lodo 131 tratados com 800 µCi, 2 em 7 tumores(29%) mostraram completa remissao sem recidiva durante o seguimento (> 3 meses). Nos mesmos casos, a utilizacao de quimioterapia com doxorubicina, 5 fluor-uracil, cisplatinum e methotrexate exibiram uma resposta menor, demonstrando um efeito antitumoral mais fugaz, nao ocorrendo nenhuma cura com o emprego de antiblasticos. Este dados sugerem que o emprego da radio-imunoterapia no tratamento do câncer da cabeca e pescoco, pode se transformar numa nova alternativa no tratamento das lesoes com altas capacidade de recidiva e disseminacao.

Monoclonal Antibodies for Diagnosis and Treatment

One of the marvels of the host immune response is its response to antigenic foreign substances by manufacturing proteins that bind tenaciously to their targets. These proteins are antibodies or immunoglobulins produced in vast diversity during an individual's lifetime. By virtue of this process, the mammalian host possesses the innate ability to mount an initial response to antigens to which there has been no prior experience and to develop an even more effective response on reexposure to these same substances. This capacity to distinguish self from nonself is one of the most basic aspects of the cellular and humoral arms of the immune response and is one of the primary means by which the host combats infection caused by many different types of pathogens. In this context, antibodies have long been recognized as a critical component of host defenses and are capable of binding to invading microbes and microbial toxins.

PATHOGENESIS AND PATHOLOGY

Pathogenesis consists of a discussion of the role of oncogenes and suppressor genes on small-cell lung cancer and non-small-cell lung cancer as well as the external factors of smoking (active and passive), asbestos, and radon. Pathology consists of discussion of squamous cell lung cancer, adenocarcinoma and bronchoalveolar carcinoma, large cell carcinoma (including giant cell and clear cell variants), and neuroendocrine tumors. Mesotheliomas are discussed as is the role of monoclonal antibodies in diagnosis.

Monoclonal Antibodies for Diagnosis and Potential Therapy of Small Cell Lung Cancer—The Ganglioside Antigen Fucosyl-Gm1

Lung carcinomas represent a highly heterogenous group of tumors, which includes small cell lung cancer (SCLC). The ganglioside fucosyl-GM1 (FucGM1) was originally identified as a highly selective marker for SCLC. A number of monoclonal antibodies against FucGM1 have been produced against the purified ganglioside, and their specificity validated. With monoclonal antibodies, FucGM1 has been detected in tissues as well as in serum samples from SCLC patients. A sensitive, quantitative assay method for FucGM1 was developed based on scintillation proximity immunoassay (SPA): A specific monoclonal antibody against FucGM1 is bound to immunosorbent particles that contain a fluor. When radiolabelled FucGM1 binds to these particles, the fluor is excited, and the photons emitted can be measured directly in a beta-counter. This sensitive assay for FucGM1 has several potential diagnostic applications: differential diagnosis of SCLC, development of serum assays for diagnosis and monitoring of disease progression, and monitoring of patient response to therapy. The expression of FucGM1 in SCLC cells is heterogeneous, and may depend on the differentiated state of tumor cells. However, monoclonal antibodies specific for FucGM1 may have important future applications for radioimmunoimaging as well as in immunotherapy for SCLC.

Monoclonal Antibodies in Diagnosis and Therapy

Monoclonal antibodies have been applied clinically to the diagnosis and therapy of an array of human disorders, including cancer and infectious diseases, and have been used for the modulation of immune responses. Effective therapy using unmodified monoclonal antibodies has, however, been elusive. Recently, monoclonal antibody-mediated therapy has been revolutionized by advances such as the definition of cell-surface structures on abnormal cells as targets for effective monoclonal antibody action, genetic engineering to create less immunogenic and more effective monoclonal antibodies, and the arming of such antibodies with toxins or radionuclides to enhance their effector function.

Role of monoclonal antibody therapy in the treatment of infectious disease

The past, present, and emerging roles of immunotherapy, including the use of monoclonal antibodies for diagnosis and treatment, are discussed. Although immunotherapy has been used for more than 100 years, it became less important when antimicrobial agents came into widespread use. In the 1970s investigators began to re-examine immunotherapy for potential use in gram-negative infections. Polyclonal antiserum against the J5 mutant of Escherichia coli (gram-negative lipid A) has been shown to be effective in treating patients with bacteremia and septic shock. The discovery of monoclonal antibodies and the creation of hybridoma technology by the fusion of immortal cells with antibody-producing cells have resulted in the production of large amounts of monoclonal antibodies of desired specificities. More recently, murine monoclonal antibodies have been used clinically for immunosuppression in renal-transplant patients (OKT3 antibody) and for prevention of septic complications in patients with suspected gram-negative infection and evidence of systemic response (E5 IgM antibody). E5 antibody directed against gram-negative bacterial endotoxin has been reported to significantly reduce morbidity and mortality from gram-negative sepsis and to be well tolerated. The application of new treatment modalities such as monoclonal antibodies is expected to enhance the therapeutic options available to treat infectious diseases.

MY7 Monoclonal Antibody for Diagnosis of Cutaneous T-Cell Lymphoma

Infiltrate in cutaneous T-cell lymphomas (CTCLs) is composed mainly of CD4 helper cells with a phenotype very similar to that of benign cutaneous lymphoid infiltrate. MY7 (CD13) is a monoclonal antibody that is normally expressed on peripheral granulocytes and monocytes but also cross-reacts with an antigen expressed on epidermal basal cells. We studied MY7 expression on basal cells of the epidermis and CD4 cell infiltrate in 34 CTCLs, 11 pseudolymphomas, and 29 other benign cutaneous lesions. An indirect immunofluorescence technique with double labeling and an immunoperoxidase technique were used. We found that in benign inflammatory infiltrate, less than 10% of CD4 cells expressed MY7 antigen associated with normal MY7 monoclonal antibody labeling of basal cells, whereas in CTCLs more than 50% of CD4 tumoral cells in dermis expressed MY7 antigens; however, basal cells were MY7 negative. Thus, it is demonstrated that MY7 monoclonal antibody with its double modulation on epidermis (basal cells) and dermis (CD4 cells) has diagnostic value for differentiating CTCLs with CD4+ MY7+ tumor cells in dermis and MY7-negative basal cells from benign inflammatory lesions with CD4+ MY7- cells in dermis and MY7-positive basal cells. This modulation of MY7 labeling could be related to the secretion of epidermal cytokines.

Application of Monoclonal Antibodies in Diagnosis of Human Breast Cancer by Fine-Needle Aspiration Cytology

In the present study the authors have examined 100 breast nodules by fine-needle aspiration cytology (FNAC) using the avidin-biotin complex method of immunoperoxidase staining. The monoclonal antibodies (MAbs) employed, B 6.2 and B 72.3, recognize two distinct molecules expressed by transformed breast epithelium. The authors have compared this immunocytochemical reaction with the conventional cytology and the histological examination as well. The data of the MAbs alone or in combination have been evaluated in terms of sensitivity, specificity, predictive value and test efficiency. The results show that tumor cells are strongly reactive with the MAbs. So the authors believe that this method in conjunction with conventional cytology may be useful in the diagnosis of breast tumor, especially in doubtful FNAC.

Second international workshop on monoclonal antibodies and breast cancer. San Francisco, November 20-21, 1986.

The Second International Workshop on Monoclonal Antibodies and Breast Cancer, organized by the John Muir Cancer and Aging Research Institute*, and chaired by Dr. Roberto L. Ceriani, brought together nearly 200 scientists, health professionals and corporate officers of 12 different countries, with an active interest in the development and application of monoclonal antibody techniques in breast cancer. In the past several years, monoclonal antibodies (MCAs) have become very powerful research tools in the biomedical sciences, and more and more laboratories are utilizing MCAs to study experimental and clinical breast cancer. The First Workshop, in 1984, also organized by the John Muir Cancer and Aging Research Institute, already suggested that MCAs might play an important role in the diagnosis and treatment of breast cancer. Since then, many more studies have been conducted to determine the benefits and limitations of MCAs in clinical practice. This review covers the 23 presentations given at the plenary sessions. They comprise a wide range of MCA research and may be divided into four main areas: characterization of antigens in normal and malignant breast tissue; evaluation of steroid receptor status in breast cancer using MCAs; technical advances and problems in the production and application of MCAs; and clinical applications of MCAs in diagnosis and treatment. In addition, 42 posters were presented which provided up-to-date information on current projects in many laboratories around the world; the posters will not be summarized here, Characterization of antigens in normal and malignant breast tissue

Monoclonal Antibodies for Diagnosis and Research in Enteric Nervous System Pathology

Monoclonal antibodies (MAb) directed against neuron-specific epitopes are valuable tools in the diagnosis of congenital and acquired enteric nervous system anomalies. MAb raised against cytoskeleton proteins (neurofilaments) revealed a characteristic staining pattern in patients with various motility disorders of the gut. Application of MAb in the study of the development of the enteric nervous system in the chicken embryo provided new insights into the fate of migrating neural crest cells. The relationship between mesenchymal target cells in the gut and proliferating neural crest cells was studied by means of MAb raised against cell surface markers (HNK-1) in combination with characterization of the microenvironment using monoclonal antibodies raised against cell adhesion molecules (N-CAM).

Use of monoclonal antibodies in the indirect fluorescent antibody technique (IFA) for the diagnosis of Edwardsiella ictaluri*

Abstract. Forty‐five channel catfish, Ictalurus pitnctaius (Rafinesque), fingerlings were inoculated with Edwardsiella ictaluri to determine the usefulness of monoclonal antibodies for indirect fluorescent antibody techniques for confirming clinical diagnosis of enteric septicaemia of catfish. Bacteriological and indirect fluorescent antibody results were compared statistically and found to correlate in 90·3% of the cases. A method for incorporating monoclonals for indirect fluorescent antibody into the speciation of Edwardsiella ictaluri and the use of monoclonal antibodies in diagnosis is described.