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Biomedical subjects

M E Carver

Publications and source records attributed to M E Carver.

11 recordsLinked to original sources
PubMed1992

Humanization of an anti-p185HER2 antibody for human cancer therapy.

The murine monoclonal antibody mumAb4D5, directed against human epidermal growth factor receptor 2 (p185HER2), specifically inhibits proliferation of human tumor cells overexpressing p185HER2. However, the efficacy of mumAb4D5 in human cancer therapy is likely to be limited by a human anti-mouse antibody response and lack of effector functions. A "humanized" antibody, humAb4D5-1, containing only the antigen binding loops from mumAb4D5 and human variable region framework residues plus IgG1 constant domains was constructed. Light- and heavy-chain variable regions were simultaneously humanized in one step by "gene conversion mutagenesis" using 311-mer and 361-mer preassembled oligonucleotides, respectively. The humAb4D5-1 variant does not block the proliferation of human breast carcinoma SK-BR-3 cells, which overexpress p185HER2, despite tight antigen binding (Kd = 25 nM). One of seven additional humanized variants designed by molecular modeling (humAb4D5-8) binds the p185HER2 antigen 250-fold and 3-fold more tightly than humAb4D5-1 and mumAb4D5, respectively. In addition, humAb4D5-8 has potency comparable to the murine antibody in blocking SK-BR-3 cell proliferation. Furthermore, humAb4D5-8 is much more efficient in supporting antibody-dependent cellular cytotoxicity against SK-BR-3 cells than mumAb4D5, but it does not efficiently kill WI-38 cells, which express p185HER2 at lower levels.

Adenocarcinoma↗
PubMed1992

High level Escherichia coli expression and production of a bivalent humanized antibody fragment.

Many clinical uses of antibodies will require large quantities of fragments which are bivalent and humanized. We therefore attempted to generate humanized F(ab')2 fragments by secretion from E. coli. Titers of 1-2 g l-1 of soluble and functional Fab' fragments have been routinely achieved as judged by antigen-binding ELISA. Surprisingly, this high expression level of Fab' in the periplasmic space of E. coli does not drive dimerization. However, we have developed a protocol to directly and efficiently recover Fab' with the single hinge cysteine in the free thiol state, allowing F(ab')2 formation by chemically-directed coupling in vitro. The E. coli derived humanized F(ab')2 fragment is indistinguishable from F(ab')2 derived from limited proteolysis of intact antibody in its binding affinity for the antigen, p185HER2, and anti-proliferative activity against the human breast tumor cell line, SK-BR-3, which over-expresses p185HER2. This system makes E. coli expression of bivalent antibody fragments for human therapy (or other uses) practical.

Amino Acid Sequence↗
PubMed1990

The extracellular domain of HER2/neu is a potential immunogen for active specific immunotherapy of breast cancer.

The proto-oncogene HER2/neu encodes a protein tyrosine kinase (p185HER2) that is homologous to the human epidermal growth factor receptor. Amplification and/or overexpression of HER2/neu occurs in multiple human malignancies and appears to be integrally involved in progression of some breast and ovarian cancers. Because of this fact, HER2/neu is an intriguing target for specific cancer therapeutic strategies. One such strategy is active specific immunotherapy, in which the immune system is targeted at specific antigens expressed by tumor cells. We have employed a transfected cell line that secretes the extracellular domain of p185HER2 as a source of HER2-derived immunogen in a guinea pig model. The immunized animals developed a cellular immune response, as monitored by delayed-type hypersensitivity, and antisera derived from immunized animals specifically inhibited the in vitro growth of human breast tumor cells overexpressing p185HER2. These data provide support for an immunotherapeutic approach to cancers characterized by overexpression of the HER2/neu proto-oncogene.

Animals↗
PubMed1986

Serum-free in vitro bioassay for the detection of tumor necrosis factor.

A sensitive, reproducible in vitro bioassay is described for quantitating the cytolytic activity of tumor necrosis factor (TNF). The assay target cells, murine connective tissue L-M, are propagated and the assay performed under serum-free conditions. The quantitation of cytolytic activity is based on the ability of TNF to lyse L-M cells in the presence of actinomycin D, as measured by crystal violet dye uptake of residual viable cells. The assay is sensitive to 88 pg/ml TNF-alpha. The simplicity of the culture medium combined with high sensitivity and low variability make this a particularly well-suited bioassay for routine detection of TNF cytolytic activity.

Biological Assay↗
PubMed1986

Comparison of TNF-alpha and TNF-beta cytolytic biological activities in a serum-free bioassay.

The effect of temperature and serum-free conditions on the cytolytic biological activities of TNF-alpha and TNF-beta were evaluated using murine connective tissue L-M as the target cell line. Serum-free assay medium and a 39 degrees C incubation temperature substantially enhanced the activity of both cytokines. L-M cells were 3 to 4 times more sensitive to cytolysis by TNF-beta than TNF-alpha under serum-free assay conditions.

Animals↗
PubMed1979

Interpretation of serum total calcium: effects of adjustment for albumin concentration on frequency of abnormal values and on detection of change in the individual.

Serum total calcium was measured in 1693 patients during a four-month period. We examined the effects of adjustment for albumin concentration on the interpretation of single measurements of serum total calcium and on the variation of series of measurements in individual patients. Markedly abnormal total calcium concentrations--2.75 mmol/l (11.0 mg/100 ml) or more, or 2.00 mmol/l (8.0 mg/100 ml) or less--were found in 115 patients, but only 24 (21%) remained markedly abnormal after adjustment for albumin. Three patients, two with malignant disease and one with primary hyperparathyroidism, had significant hypercalcaemia which was masked by hypoalbuminaemia. The serum total calcium measured on a subsequent occasion had changed 0.15 mmol/l (0.6 mg/100 ml) or more in 60 patients, but after adjustment for albumin this number was reduced to 27 (45%). The within-person standard deviation for serum total calcium was calculated in 26 patients with normal mean adjusted calcium concentrations who had had six or more sequential measurements. The mean standard deviation was 0.148 mmol/1 (0.59 mg/100 ml) and, after adjustment for albumin, this was reduced to 0.100 mmol/1 (0.40 mg/100 ml). We conclude that adjustment of serum total calcium concentration for albumin is essential to detect abnormal values and to assess changes in a value.

Blood Protein Disorders↗
PubMed1977

Plasma creatinine and urea: creatinine ratio in patients with raised plasma urea.

We examined the plasma urea and creatinine concentrations and the ratio between them according to diagnosis in 100 unselected and 31 selected adult hospital patients with a plasma urea concentration greater than or equal to 10 mmol/l (60mg/100ml). We also examined plasma urea and creatinine concentrations in 350 unselected consecutive patients, but found no useful relation between the two values. Congestive heart failure was the most common identifiable cause of a raised plasma urea concentration in the 100 unselected patients (36%). Among these 100 patinets the plasma creatinine concentration was a more useful discriminant between prerenal uraemia and intrinsic renal failure than was the urea:creatinine ratio or the plasma urea concentration. A plasma creatinine concentration greater than 250 mumol/1 (2-8 mg/100ml) indicated intrinsic renal failure with a 90% probability.

Acute Kidney Injury↗
PubMed1977

Maternal nutrition in early pregnancy.

1. Mean daily nutrient intakes of 195 women in the first trimester of pregnancy were assessed by weighed dietary records. 2. In comparsion with recommended intakes for non-pregnant women aged 18-55 years (Department of Health and Social Security, 1969), more than two-thirds of the subjects were having insufficient energy, iron and cholecalciferol. Unsatisfactory intakes of other nutrients were not uncommon. In relation to recommended intakes for the second trimester (Department of Health and Social Security, 1969), all mothers were having insufficient cholecalciferol and more than 80% of mothers had unsatisfactory intakes of energy and Fe. 3. Intakes appreciably lower than those recommended were associated with the following factors: social classes III, IV and V; maternal age under 20 years; smoking ten or more cigarettes daily; vomiting on more than 3 d/week.

Adult↗
PubMedDate not supplied

Developing a strategy for managing the hospital's growing technology.

Successful hospital CEOs, administrators and department heads skilled at their professions, face a major challenge from the onslaught of rapidly developing medical instrumentation technology. To meet this rapidly growing challenge, they must identify their needs, survey their resources, and take positive action now to develop the necessary clinical engineering and equipment control programs to cope cost effectively with the future. This article provides an overview of the fundamental problem for the hospital administrator, suggests a direction toward solution, and provides a primer on the resources available to effectively deal with, if not solve, the enormous problem posed by the ever increasing technology dependence of health care.

Biomedical Engineering↗