Development of monoclonal antibody-coated immunomagnetic beads for separation and detection of norovirus (genogroup II) in faecal extract samples

ABSTRACT Bacteroides fragilis is a constituent of the normal resident microbiota of the human intestine and is the gram-negative obligately anaerobic bacterium most frequently isolated from clinical infection. Surface polysaccharides are implicated as potential virulence determinants. We present evidence of within strain immunochemical variation of surface polysaccharides in populations that are noncapsulate by light microscopy as determined by monoclonal antibody labelling. Expression of individual epitopes can be enriched from a population of an individual strain by use of immunomagnetic beads. Also, individual colonies in which either >94% or <7% of the bacteria carry an individual epitope retain this level of expression when subcultured into broth. In broth cultures where >94% of the bacteria carry a given epitope, there is no enrichment for other epitopes recognized by different polysaccharide-specific monoclonal antibodies. This intrastrain variation has important implications for the development of potential vaccines or immunodiagnostic tests.

Download Full-text

Diagnosis of Mediterranean Spotted Fever by Indirect Immunofluorescence of Rickettsia conorii in Circulating Endothelial Cells Isolated with Monoclonal Antibody-Coated Immunomagnetic Beads

The Journal of Infectious Diseases ◽

10.1093/infdis/166.3.660 ◽

1992 ◽

Vol 166 (3) ◽

pp. 660-663 ◽

Cited By ~ 56

Author(s):

M. Drancourt ◽

F. George ◽

P. Brouqui ◽

J. Sampol ◽

D. Raoult

Keyword(s):

Monoclonal Antibody ◽

Endothelial Cells ◽

Indirect Immunofluorescence ◽

Spotted Fever ◽

Circulating Endothelial Cells ◽

Rickettsia Conorii ◽

Mediterranean Spotted Fever ◽

Immunomagnetic Beads

Download Full-text

Improved Recovery of Exfoliated Colonocytes from Feces Using Newly Developed Immunomagnetic Beads

Gastroenterology Research and Practice ◽

10.1155/2008/605273 ◽

2008 ◽

Vol 2008 ◽

pp. 1-7 ◽

Cited By ~ 7

Author(s):

Yoshikatsu Koga ◽

Masahiro Yasunaga ◽

Satoshi Katayose ◽

Yoshihiro Moriya ◽

Takayuki Akasu ◽

...

Keyword(s):

Colorectal Cancer ◽

Monoclonal Antibody ◽

Monoclonal Antibodies ◽

Recovery Rate ◽

Magnetic Beads ◽

Clinical Samples ◽

Immunomagnetic Beads ◽

Colorectal Cancer Cells ◽

Efficient Recovery ◽

Ht 29

We demonstrated the feasibility of a new methodology for isolating colonocytes from feces. To reduce costs and improve the recovery rate of colonocytes from feces, we attempted to develop new immunomagnetic beads. Several sizes of magnetic beads were prepared and tagged with a monoclonal antibody against EpCAM. We made several new monoclonal antibodies against EpCAM, and each monoclonal antibody was tagged to the magnetic beads. In the simulation, the most efficient recovery of HT-29 cells was obtained using the smallest size of beads. Also, beads tagged with a monoclonal antibody with a higher affinity against EpCAM had a higher recovery rate. Similar results were obtained when the smallest size of beads with the highest-affinity monoclonal antibody was applied to clinical samples. The newly developed immunomagnetic beads may be useful for isolating colorectal cancer cells from feces, enabling the cytological or molecular biological diagnosis of CRC.

Download Full-text

Development of A Monoclonal Antibody-Coated Immunomagnetic Beads for Separation and Detection of Norovirus (Genogroup II) in Oysters

Food Science and Technology Research ◽

10.3136/fstr.19.669 ◽

2013 ◽

Vol 19 (4) ◽

pp. 669-674

Author(s):

Lin YAO ◽

Yanhua JIANG ◽

Wei JIANG ◽

Fengling LI ◽

Yuxiu ZHAI ◽

...

Keyword(s):

Monoclonal Antibody ◽

Immunomagnetic Beads

Download Full-text

A web-like network of type vi collagen in human skin is revealed by immuno-electron microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100103978 ◽

1988 ◽

Vol 46 ◽

pp. 382-383

Author(s):

Douglas R. Keene ◽

Robert W. Glanville ◽

Eva Engvall

Keyword(s):

Electron Microscopy ◽

Monoclonal Antibody ◽

Human Skin ◽

Basement Membranes ◽

Primary Antibody ◽

Fat Cells ◽

Secondary Antibody ◽

Type Vi Collagen ◽

Dermal Matrix ◽

Immuno Electron Microscopy

A mouse monoclonal antibody (5C6) prepared against human type VI collagen (1) has been used in this study to immunolocalize type VI collagen in human skin. The enbloc method used involves exposing whole tissue pieces to primary antibody and 5 nm gold conjugated secondary antibody before fixation, and has been described in detail elsewhere (2).Biopsies were taken from individuals ranging in age from neonate to 65 years old. By immuno-electron microscopy, type VI collagen is found to be distributed as a fine branching network closely associated with (but not attached to) banded collagen fibrils containing types I and III collagen (Fig. 1). It appears to enwrap fibers, to weave between individual fibrils within a fiber, and to span the distance separating fibers, creating a “web-like network” which entraps fibers within deep papillary and reticular dermal layers (Fig. 2). Relative to that in the dermal matrix, the concentration of type VI collagen is higher around endothelial basement membranes limiting the outer boundaries of nerves, capillaries, and fat cells (Fig. 3).

Download Full-text

High-voltage electron microscopy of subretinal scar formation

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100122836 ◽

1992 ◽

Vol 50 (1) ◽

pp. 486-487

Author(s):

G.E. Korte ◽

M. Marko ◽

G. Hageman

Keyword(s):

Electron Microscopy ◽

Monoclonal Antibody ◽

Retinal Pigment Epithelium ◽

Glial Cells ◽

High Voltage ◽

Pigment Epithelium ◽

Retinal Pigment ◽

Sodium Iodate ◽

High Voltage Electron Microscopy ◽

Voltage Electron

Sodium iodate iv. damages the retinal pigment epithelium (RPE) in rabbits. Where RPE does not regenerate (e.g., 1,2) Muller glial cells (MC) forma subretinal scar that replaces RPE. The MC response was studied by HVEM in 3D computer reconstructions of serial thick sections, made using the STEREC0N program (3), and the HVEM at the NYS Dept. of Health in Albany, NY. Tissue was processed for HVEM or immunofluorescence localization of a monoclonal antibody recognizing MG microvilli (4).

Download Full-text

Identification of the pulmonary syndrome hantavirus by direct and colloidal gold immune Electron Microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100169109 ◽

1994 ◽

Vol 52 ◽

pp. 274-275

Author(s):

C. D. Humphrey ◽

C.S. Goldsmith ◽

L. Elliott ◽

S.R. Zaki

Keyword(s):

Electron Microscopy ◽

United States ◽

Monoclonal Antibody ◽

Colloidal Gold ◽

Phosphotungstic Acid ◽

Uranyl Acetate ◽

Hantavirus Pulmonary Syndrome ◽

Deer Mice ◽

Immune Electron Microscopy ◽

Negative Stain

An outbreak of unexplained acute pulmonary syndrome with high fatality was recognized in the spring of 1993 in the southwestern United States. The cause of the illness was quickly identified serologically and genetically as a hantavirus and the disease was named hantavirus pulmonary syndrome (HPS). Recently, the virus was isolated from deer mice which had been trapped near the homes of HPS patients, and cultivated in Vero E6 cells. We identified the cultivated virus by negative-stain direct and colloidal gold immune electron microscopy (EM).Virus was extracted, clarified, and concentrated from unfixed and 0.25% glutaraldehyde fixed supernatant fluids of infected Vero E6 cells by a procedure described previously. Concentrated virus suspensions tested by direct EM were applied to glow-discharge treated formvar-carbon filmed grids, blotted, and stained with 0.5% uranyl acetate (UA) or with 2% phosphotungstic acid (PTA) pH 6.5. Virus suspensions for immune colloidal gold identification were adsorbed similarly to filmed grids but incubated for 1 hr on drops of 1:50 diluted monoclonal antibody to Prospect Hill virus nucleoprotein or with 1:50 diluted sera from HPS virus infected deer mice.

Download Full-text