Use Of Monoclonal Antibodies Research Articles

Aptamers in Therapeutics

Aptamers are single strand DNA or RNA molecules, selected by an iterative process known as Systematic Evolution of Ligands by Exponential Enrichment (SELEX). Due to various advantages of aptamers such as high temperature stability, animal free, cost effective production and its high affinity and selectivity for its target make them attractive alternatives to monoclonal antibody for use in diagnostic and therapeutic purposes. Aptamer has been generated against vesicular endothelial growth factor 165 involved in age related macular degeneracy. Macugen was the first FDA approved aptamer based drug that was commercialized. Later other aptamers were also developed against blood clotting proteins, cancer proteins, antibody E, agents involved in diabetes nephropathy, autoantibodies involved in autoimmune disorders, etc. Aptamers have also been developed against viruses and could work with other antiviral agents in treating infections.

Minimal tumour burden in haematological diseases: a step forward with quantitative assessment of Bence-Jones in nephelometry?

Minimal tumour burden in haematological diseases: a step forward with quantitative assessment of Bence-Jones in nephelometry?

Structural Characterization of Nanoyeast Single-Chain Fragment Variable Affinity Reagents

Nanoyeast single-chain variable fragments (nanoyeast-scFv) are a new class of low-cost and stable protein capture agents developed as alternatives to full length monoclonal antibodies for use in immunosensors. Physical characteristics that impart nanoyeast-scFv with these advantages have yet to be investigated. We investigate the structure, size and surface loading of nanoyeast-scFv to better understand its ability to specifically and sensitively capture proteins of interest while retaining activity in solution. Nanoyeast-scFv fragments were found to be globular in structure and heterogeneous in size, typically <100 nm, and were found to aggregate in solution. Optimal fragment size was determined to be <220 nm, with larger fragments interfering with the electrochemical biosensor. We hypothesize that the stability of nanoyeast-scFv is conferred in part by components of the yeast cell wall fragments, which provide stability to the associated scFv both during selection, lyophilization, and rehydration.

Design and in Silico Construction of New Efficient Antibodies Against Gastric Tumor-associated Antigens Towards Introducing Combined Strategy for Vaccine Tumor Therapy

Cancer with more than 11 million deaths per year is the second principal cause of death. It is estimated that there will be 16 million new cases every year by 2020. Deaths from cancer are predicted to be rising, with an expected 9 million people dying from cancer in 2015 and 11.4 million dying in 2030, worldwide. Additionally, gastric cancer is the second most common form of cancer-related deaths, globally. Cancer immunotherapy has been introduced as an influential and promising clinical trend for the treatment of cancer and it has been successful in treatment of breast and prostate cancers as well as melanoma [1]. Tumors are appeared by the progressive growth and proliferation of non-programmed cells. It is probable that cancerous cells occur daily in healthy individuals; however in the majority of cases, they are detached by the immune system and cannot grow into malignancies. The above-mentioned capability of the immune system to sense and eliminate cancerous cells is known as “immune surveillance”. Cancerous cells are detected by innate and adaptive immune cells which sense the local environment of a malignancy when it begins to remodel the stroma [1]. The crucial aim of immunotherapy is overcoming malignancies by inducing an efficient immune response against the cancerous cells. In theory, if particular antigens could be recognized in the forerunner lesions leading to a tumor, the immune responses can be used to eradicate them which may prevent the cancerous cells to convert into a malignancy [2]. Gastric cancer is potentially a suitable target for such defensive immunotherapy as it progresses over an extended period of time and its premalignant lesions are clearly identifiable which if effectively targeted, their development to forthright tumor could be prevented. Current immune therapies of gastric tumors are frequently based on utilizing monoclonal antibodies (vaccine and drugs), cytotoxic immunocytes, or gene transferred vaccines [3]. Experimental vaccination methodologies involve using whole protein and peptide vaccines, based on recognition of certain peptides by cytotoxic ABSTRACT Introduction: Gastric cancer, as the second reason of cancer mortality around the world, is defined as one of the serious health problems and a wide range of current research is focused to overcome the challenges relevant to its successful therapy. Employing recent approaches in the field of vaccine tumor therapy and immunotherapic strategies such as utilization of monoclonal antibodies can play significant roles in achieving efficient therapeutic means to combat the disease. Methods: In this study, computational methods including three-dimensional modeling, based on sequence combinational pattern identification together with docking procedures have been employed for design and in silico construction of prolific and effective antibody structures against HER-2 and MAGE-3 antigens. Results: These methods led to introduction of novel and efficient anti-HER2 and anti-MAGE3 antibodies. For instance, combinational pattern of bH1 antibody (light chain) and IGG1-KAPPA 4D5 antibody (heavy chain) which obtained an exceptional score of 844.83 KJ/mol as the new candidate for cancer vaccine therapy, whereas Herceptin scored 453.4 KJ/mol. In addition, the suggested pattern for designing antibody against MAGE-3 with combination of bH1 antibody (heavy chain) and HLA-A 0201 (light chain) achieved acceptable score of 640.8 KJ/mol. Conclusion: It is envisaged that the results obtained from the current research could be utilized to develop efficient combined strategies for vaccine therapy of gastric cancer.

Antibodies and their multivalent constructs for cancer therapy.

In 2008 cancer was identified by the World Health Organization (WHO) as one of four threats to human health and development. Since the early published reports of the first chemotherapeutic, mustine, in 1946, the anti-cancer drug and development industry has grown into a multi-billion dollar business enterprise. Worldwide, the rates of new cancer cases and deaths has been steadily increasing each year, with the estimation by the WHO-sponsored GLOBOCAN cancer database, that at current rates, nearly 13 million cancer deaths will be reported in 2030. The recent successes of monoclonal antibodies (mAbs), an important class of glycoprotein, and their multivalent and drug conjugated derivatives over the past 30 years have led to the approval of 12 monoclonal antibodies for use in cancer treatment by the FDA. Modern recombinant and engineering techniques have led to an explosion of antibody platforms that can be attributed to great gains in clinical efficacy. This review discusses and outlines a sample of mAbs currently approved for cancer treatment by the FDA, as well as antibody platforms in the research pipeline and clinic that have been engineered for greater tumor penetration, binding, and efficacy.

Cancer immunotherapeutics: evolution of monoclonal antibodies to peptide immunogens.

The Wistar Institute under Hilary Koprowski's direction played a preeminent role in bringing the fruits of basic science to clinic through biotechnology and life science enterprises. Koprowski's early view on the utility of monoclonal antibodies has been validated to some extent, because monoclonal therapeutics form one of the fastest growing and most successful and lucrative segments within the biopharmaceutical sector. Over 30 monoclonal antibody drugs have now been approved in the United States and Europe. However, monoclonal antibodies might be viewed in the context of a bridge to the future for vaccines. The Wistar Institute's reputation, while built on vaccines for infectious diseases, can be translated in years to come into vaccines for cancer.

Production of recombinant antibodies using bacteriophages.

Recombinant antibody fragments such as Fab, scFv, diabodies, triabodies, single domain antibodies and minibodies have recently emerged as potential alternatives to monoclonal antibodies, which can be engineered using phage display technology. These antibodies match the strengths of conventionally produced monoclonal antibodies and offer advantages for the development of immunodiagnostic kits and assays. These fragments not only retain the specificity of the whole monoclonal antibodies but also easy to express and produce in prokaryotic expression system. Further, these antibody fragments are genetically stable, less expensive, easy to modify in response to viral mutations and safer than monoclonal antibodies for use in diagnostic and therapeutic applications. This review describes the potential of antibody fragments generated using phage display and their use as diagnostic reagents.

Antibody Discovery from Immune Competent and Immune Transplanted Mice

Since Kohler and Milstein developed the process of generating hybridomas by fusing antibody secreting B cells with an immortal myeloma cell line, the techniques used to develop monoclonal antibodies for use as human therapeutics have progressed significantly. Here, we will briefly review hybridoma technology and the evolution of therapeutic antibodies for the treatment of human disease. We will focus on the evolution of humanized mouse models for the generation of therapeutic human antibodies, comparing the early models, such as severe combined immunodeficient (SCID) mice which do not engraft human leukocytes well due to residual innate immunity, to the more recently developed models such as non-obese diabetic (NOD)/SCID IL-2Rγ-deficient mice in which numerous human hematopoietic lineages can be cultivated. Building on the identification of suitable host strains for the reconstitution of human immune cells, focus has now shifted onto humanizing the murine microenvironment in order to support human immune cell function. Although several recent studies have shown that the provision of human soluble factors can support maturation and function of human immune cells, particularly within the myeloid compartment, this does not appear to impact antibody production significantly. Moreover, models in which grafting of human tissues is performed to provide human microenvironments which support human leukocyte maturation do show improved humoral immune function, but require several surgical manipulations for generation of the model. Ultimately the most desirable scenario is to generate transgenic models that can be bred efficiently and express a sufficient number of human molecules to support functional human immune cells and several groups have made progress in making this idea a reality. These studies in the context of the generation of human antibodies will be discussed.

Immunotherapy for Gastrointestinal Malignancies

Gastrointestinal (GI) malignancies (esophageal, gastric, pancreatic, intra- and extra-biliary ductal, hepatocellular, and colorectal cancers) are an important cause of cancer incidence and mortality in the US and globally. GI cancers account for 15.4% and 23.8% of incident cancers and cancer-related deaths respectively in the US alone. Although earlier diagnosis and treatment advances have improved outcomes for some GI malignancies, the need for improved therapies in all disease phases (adjuvant, neoadjuvant and advanced) is paramount. Utilization of monoclonal antibodies targeting against vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR) has shown the success in selected colorectal carcinoma patients. More investigations of immunotherapy are on going in the treatment of GI malignances with different mechanisms and methods. In this article, we review data for established and evolving immunotherapy-related treatment options in GI malignancies.

SP0124 Infections and rheumatic diseases: What is new?

There has been a long-standing belief that both rheumatoid arthritis and other types of inflammatory arthritis may be a consequence, either directly or indirectly, of infection. Although there is no...

Characterization of a human CD27 agonistic monoclonal antibody for use as an agent in glioblastoma therapy (P4411)

Abstract CD27 is a costimulatory molecule constitutively expressed on naïve T-cells, B-cells, and subsets of NK cells; signaling through CD27 has been shown to promote Th1 polarization, clonal expansion, enhanced survival of effector and memory cells, upregulation of effector molecules, and enhanced cytolytic activity. We are characterizing the use of an agonistic anti-CD27 antibody, CDX1127, as a treatment in a preclinical glioblastoma model in human CD27 transgenic mice. We have shown that in vitro stimulation of isolated T-cells from hCD27+/- mice with CDX1127, in the context of TCR signaling, leads to increased proliferation, decreased apoptosis, and increased secretion of IFN-γ and TNF-α, corroborating its agonistic activity. Furthermore, we have analyzed the efficacy of CDX1127 as a monotherapy in a mouse glioblastoma model as well as an adjuvant in a total tumor RNA dendritic cell vaccine.

A Monoclonal Antibody Against Bovine Adiponectin

Adiponectin (AdipoQ) is an adipokine mainly secreted by white fatty tissue, playing a major role in energy homeostasis and insulin sensitivity. For cattle, AdipoQ data are largely limited to mRNA expression; to our knowledge, valid information about the AdipoQ protein in bovine tissues and body fluids is not available. Therefore, we have developed a monoclonal antibody against bovine AdipoQ. This study describes the preparation, application, and characterization of a monoclonal antibody for use in ELISA, Western blot, and histology. The antibody was developed by PEG fusion of the SP2/0 cell line with splenic B cells from AdipoQ immunized C57Bl/6 mice. Antibody-producing cells were identified by ELISA and specified by immunoblotting and immunostaining of bovine retroperitoneal adipose tissue. The novel antibody detects AdipoQ in histological samples, ELISA, and Western blots.

Characterization and utility of monoclonal antibodies against spike protein of transmissible gastroenteritis virus

Aims: This work aims to characterize the utility of four newly generated monoclonal antibodies (mAbs) against transmissible gastroenteritis virus (TGEV). Methods and Results: Four monoclonal antibodies (mAbs) against the N‐terminal half of spike protein (S1 protein) of TGEV were identified. Affinity constant of these mAbs was analysed. These mAbs were capable of reacting with the TGEV S1 protein analysed by ELISA and Western blot. A competition assay between the different mAbs was performed to determine whether the different antibodies mapped in the same or a different antigenic region of the protein. Investigation on the neutralizing ability of these mAbs indicated that two of these mAbs completely neutralized TGEV at an appropriate concentration. These mAbs were able to detect the TGEV‐infected cells in immunofluorescence assays and Western blot. Moreover, they differentiated TGEV S protein from other control proteins. Conclusions: The generated four mAbs are very specific, and the established immunofluorescence assays, Western blot and discrimination ELISA are useful approaches for detecting of TGEV. Significance and Impact of the Study: It is a novel report regarding the use of the S1 protein of TGEV to generate specific mAbs. Their utility and the established immunoassays contribute to the surveillance of TGE coronavirus.

MALDI Immunoscreening (MiSCREEN): A method for selection of anti-peptide monoclonal antibodies for use in immunoproteomics

A scalable method for screening and selection of peptide-specific monoclonal antibodies (mAbs) is described. To identify high affinity anti-peptide mAbs in hybridoma supernatants, antibodies were captured by magnetic affinity beads followed by binding of specific peptides from solution. After timed washing steps, the remaining bound peptides were eluted from the beads and detected by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). This allowed measurement of monovalent interactions of peptides with single antigen binding sites on the antibodies, thus reflecting antibody affinity rather than avidity. Antibodies that were able to bind target peptides from solution phase and retain them during washing for a minimum of 10 min were identified by the strength of the appropriate m/z peptide MS signals obtained. This wash time reflects the minimum peptide dissociation time required for use of these antibodies in several current immuno-mass spectrometry assays. Kinetic analysis of antibody-peptide binding by surface plasmon resonance (SPR) showed that the selected antibodies were of high affinity and, most importantly, had low dissociation constants. This method, called MALDI immunoscreening (MiSCREEN), thus enables rapid screening and selection of high affinity anti-peptide antibodies that are useful for a variety of immunoproteomics applications. To demonstrate their functional utility in immuno-mass spectrometry assays, we used the selected, purified RabMAbs to enrich natural (tryptic) peptides from digested human plasma.

Patient Characteristics Associated with Use of Monoclonal Antibody Treatment in Women with Metastatic Breast Cancer: A Population-Based Analysis.

Abstract Introduction: Monoclonal antibodies (MoATB) were FDA approved for the treatment of patients with breast cancer (BrCA) whose tumors over expressed the human epidermal growth factor receptor 2 (HER2) in 1998. We sought to determine if certain patient characteristics and co-morbidities influenced MoATB use in a population-based practice setting of elderly women with metastatic BrCA. Methods: We identified female Medicare beneficiaries aged 66+ years diagnosed with incident metastatic BrCA without prior history for any cancer between 1999 and 2005 in the Surveillance, Epidemiology and End Results cancer registries (SEER). Charlson Co-morbidity index (CCI) and separate conditions within the index were created using linked Medicare claims in the year prior to the BrCA diagnosis. We identified both oral and infused chemotherapy (CH) from the Medicare claims from 1999 to 2006. Patients were categorized into three groups: no CH, MoATB+/-CH, and CH w/out MoATB. We performed bivariate statistics to compare the patient characteristics and comorbidities between the three groups, and the treatments groups only. Results: There were 3,820 women with metastatic BrCA, mean age 77 (SD 7.3) years, 81% were white race, 29% were married and 91% lived in an urban setting. 67% of these women (n=2,562) received no CH, 26% received CH w/out MoATB (n=994) and 7% received MoATB+/-CH (n=264). Overall the CCI was lower for women receiving either treatment as compared to no chemotherapy and between treatment groups for the young old (66-75 years) but not those aged 80+ years. The prevalence of congestive heart failure (CHF) and cerebrovascular disease was significantly lower in women receiving MoATB +/-CH, as compared to only CH, and also lower in both compared to no CH (n=2,462). Few women presenting with COPD or dementia were likely to be treated with either CH and/or MoATB compared to No CH. Other co-morbidity conditions within the CCI were not statistically different between the groups or the cell numbers were too small to analyze. Conclusion: MoATB use in the treatment of metastatic breast cancer steadily increased from 5% in 1999 to 23% in 2004. Use in 2005 was slightly lower but will be recalculated once the 2007 claims are available. In 2008, FDA released a black box warning for trastuzumab, the most frequently prescribed MoATB for BrCA, regarding the potential for the development of cardiomyopathy. Our results suggest that co-morbidities in particular CHF and cerebrovascular disease may influence the decision to use MoATB. Further research is necessary to examine the relationship between co-morbidities and the use of MoATB in the treatment of metastatic breast cancer. Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 2057.

The use of monoclonal antibodies and related agents in the treatment of respiratory disease

Monoclonal antibodies were first developed in 1975. Since their introduction they have had a progressively increasing impact on different areas of medical diagnostics and therapeutics. Currently, many monoclonal antibodies are under investigation for their potential role in pulmonary medicine. This review discusses a brief history of monoclonal antibodies; the manufacture, use, biosafety and nomenclature of these agents; the inflammatory response in asthma; and the use of omalizumab, an FDA-approved monoclonal antibody for use in asthma, will be described in detail, as well as its associated adverse effects. Additionally, other related biologic agents that may have a role in respiratory diseases are presented.

Meningoencefalite por herpesvírus bovino-5

A meningoencefalite por herpesvírus bovino-5 (BoHV-5) é uma doença infecto-contagiosa, aguda ou subaguda, geralmente fatal e que afeta principalmente bovinos jovens submetidos a situações de estresse. A doença tem sido freqüentemente diagnosticada em várias regiões do Brasil e em outras partes do mundo. BoHV-5 é um vírus da família Herpesviridae e subfamília Alphaherpesvirinae e possui como genoma uma molécula de DNA fita dupla. Esses vírus são caracterizados por rápida replicação em cultivo, que resulta em lise das células infectadas, e afetam várias espécies de hospedeiros, estabelecendo latência principalmente em neurônios de gânglios sensoriais. A transmissão de BoHV-5 ocorre principalmente por contato direto ou indireto entre bovinos. Após a replicação primária nas mucosas oral, nasal, ocular e orofaríngea, o vírus invade as terminações nervosas e é transportado até os neurônios de gânglios sensoriais, onde replica ativamente e estabelece latência. A invasão viral do encéfalo pode resultar em replicação viral massiva e produção de doença neurológica. A maioria dos bovinos que desenvolvem doença neurológica morre em decorrência de meningo-encefalite, porém alguns podem desenvolver infecção subclínica e, após recuperação, permanecerem portadores da infecção latente. A disseminação viral nos rebanhos é facilitada em situações de grande concentração de animais, introdução de bovinos e desmame de lotes de bezerros em idade que coincide com o decréscimo da imunidade passiva. Certas condições, naturais ou induzidas, podem reativar o vírus do estado latente e propiciar condições para sua transmissão e disseminação a outros indivíduos. A doença pode ocorrer na forma de surtos ou em casos isolados, com coeficientes de morbidade que podem variar de 0,05%-5%; a letalidade é quase sempre de 100%. Os sinais clínicos incluem depressão, descarga nasal e ocular, ranger de dentes, andar em círculos, cegueira, febre, movimentos de pedalagem, disfagia, dor abdominal, nistagmo, tremores, sialorréia, incoordenação, opistótono, pressão da cabeça contra objetos, quedas e convulsões. A evolução do quadro clínico pode variar de 1 a 15 dias. Achados de necropsia podem estar ausentes, mas normalmente se observa tumefação das porções rostrais do córtex telencefálico e achatamento das circunvoluções, com segmentos amarelados e amolecidos (malacia). Com a evolução da doença, essas áreas se tornam gelatinosas e acinzentadas, e em casos avançados ocorre o desaparecimento segmentar do córtex telencefálico frontal (lesão residual). Em muitos casos podem ser observados focos de malacia na substância cinzenta dos núcleos basais e do tálamo. Histologicamente observa-se meningoencefalite não-supurativa necrosante, principalmente no córtex telencefálico frontal, associada a inclusões intranucleares eosinofílicas em astrócitos e neurônios, embora a freqüência dessas inclusões seja irregular. O diagnóstico de meningoencefalite por BoHV-5 deve ser feito com base nos achados epidemiológicos, clínicos, de necropsia e histopatológicos, associados com o isolamento do vírus em cultivo celular células ou com detecção de antígenos virais em seções do encéfalo ou em células descamadas presentes nas secreções nasais. A identificação e caracterização de BoHV-5 pode ser realizada por meio de testes com anticorpos mono-clonais, reação em cadeia de polimerase (PCR) e por análise de restrição genômica. Não há tratamento específico para a meningoencefalite por BoHV-5. Como o BoHV-1 e o BoHV-5 são antigenicamente muito semelhantes, recomenda-se a vacinação com vacinas para BoHV-1 como forma de reduzir as perdas causadas por BoHV-5, principalmente durante surtos de doença neurológica. Adicionalmente, outras medidas podem ser adotadas para prevenir ou reduzir os prejuízos ocasionados pela enfermidade, como testar sorologicamente os bovinos a serem introduzidos nos rebanhos, minimizar situações de estresse, sobretudo no desmame, e isolamento dos bovinos afetados.

Progress in the discovery and definition of monoclonal antibodies for use in feline research

The practice of veterinary medicine and research into both animal diseases and animal models of human disease are restricted by the scarcity of monoclonal antibodies (mAb) that react with animal proteins. One way to enlarge the repertoire of mAb to animal leukocyte differentiation antigens (LDA) is to test mAb specific to other species for cross-reactivity to the species of interest. We have tested a panel of 380 commercially available anti-human mAb for cross-reactivity to feline LDA. Twenty-six of these mAb cross-react with cat LDA and 19 others are of questionable cross-reactivity. Definition of mAb specificity in the cat is being investigated by multi-color flow cytometry (FCM) to compare test mAb specificity with that of mAb to known feline LDA. The addition of these cross-reactive mAb to the anti-feline mAb currently available will enhance studies in comparative medicine.

Simultaneous Detection and Quantitation of Multiple Antigen Expression in Tissue by Fluorescence Microscopy and Hyper-spectral Imaging (132.4)

Abstract Analyses of biomarkers in tissues represent a critical component in the development of therapeutics. In drug development, entire signaling pathways are profiled with a variety of high-throughput techniques. However, in the field of histology, tissue-based diagnostics are generally dominated by single- or dual-analyte tests. In addition, manual methods for slide processing promote inconsistency. We have developed automated instrumentation for conducting high-throughput IHC and ISH in widely available formalin-fixed paraffin-embedded samples. In this report we describe the utilization of Quantum dots (Qdots®) in advanced antigen detection in intact tissues. Specifically, we developed a novel set of hapten and anti-hapten monoclonal antibodies for use in multi-analyte assays. Spectral imaging software was utilized to deconvolute images generated by fluorescence microscopy on tissue samples. In model systems, we demonstrate simultaneous detection, and quantitation of expression levels and cell numbers, for markers including CD20, CD34, CD45, IgGκ, IgGλ, and Ki67. These reagents provide a growing set of tools that could complement early marker validation. The examples in this study demonstrate the utility of coupling high-throughput, automated histochemistry with quantifiable, multiplexed analyte detection. These findings are reported for individual cells in their anatomic context.

Survey of select practice behaviors by primary care physicians on the use of opioids for chronic pain

ABSTRACTBackground: Severe allergic asthma patients may not be controlled even with guideline recommended care, including inhaled corticosteroids, long-acting beta-2 agonists, theophylline, oral steroids and anti-leukotrienes. They experience exacerbations requiring intensive healthcare use and which may be fatal. Omalizumab, a new monoclonal antibody for use in IgE-mediated allergic diseases, reduces exacerbations and daily symptoms in this patient population. The aim of this study is to estimate the cost effectiveness of adding omalizumab to optimized standard therapy (ST) in patients with severe persistent IgE-mediated (allergic) asthma.Objective: To assess the use of opioids by primary care physicians for the treatment of chronic pain.Methods: A written survey was completed by 248 primary care physicians. Outcomes of interest included type of opioids prescribed, common pain diagnoses treated, opioid prescribing concerns, treatment of patients with a history of substance use disorders and clinic-based protocols for pain management.Results: The mean age of the physicians who completed the questionnaire was 41 years. The majority were between the ages of 30 and 49 years (68%) with an equal number of men and women. Seventy percent were family physicians, 28.7% internists and less than 2% were community physicians and geriatricians. Physician concerns regarding opioid therapy included prescription drug abuse (84.2%), addiction (74.9%), adverse effects (68%), tolerance (60.7%), and medication interaction (32%). The survey found that the majority of the physicians were comfortable in prescribing narcotics to patients with terminal cancer. However, they were less comfortable prescribing narcotics to patients with low back pain and persons with a current or past history of drug or alcohol abuse. Physician management practices suggested that urine toxicology tests were under-utilized with only 6.9% reporting obtaining this test before prescribing opioids and only 15.0% performing urine toxicology tests on patients already prescribed opioids. Logistic regression analysis revealed that whether or not physicians routinely conducted urine toxicology screens was significantly (p = 0.015) predicted by whether they had a system to track patients on opioids when prescribing narcotics. The primary limitation of the study is the reliance on physician self-report rather than objective measures of physician behavior.Conclusions: The survey suggests physicians are concerned about drug abuse, addiction, adverse effects, tolerance, and medication interaction. Their comfort level in prescribing opioids varies with the patient characteristics. Urine toxicology testing is underutilized in the primary care setting.