Monoclonal antibodies to human A-I apolipoprotein and characterisation of cyanogen bromide fragments of APOA-I

ABSTRACT Two reference monoclonal antibodies against the meningococcal P1.15 subtype PorA, MN3C5C and 2-1-P1.15, showed only partial concordant recognition of meningococcal isolates. Cyanogen bromide cleavage of P1.19,15 PorA, peptide mapping, and sequencing of porAregions demonstrated that 2-1-P1.15 was specific for subtype P1.19, and henceforth it is to be redesignated as 2-1-P1.19.

Download Full-text

Characterization Of A Crosslink-Containing Fragment Derived From The A Polymer Of Human Fibrin And Its Application In Immunologic Studies Using Monoclonal Antibodies

10.1055/s-0038-1652710 ◽

1981 ◽

Author(s):

J H Sobel ◽

S Birken ◽

P Ehrlich ◽

R Friedman ◽

Z Moustafa ◽

...

Keyword(s):

Molecular Weight ◽

Monoclonal Antibodies ◽

Early Detection ◽

High Molecular Weight ◽

Primary Structure ◽

Cyanogen Bromide ◽

Tryptic Peptide ◽

Structural Studies ◽

Polymer Component

A high molecular weight crosslink-containing fragment derived from cyanogen bromide (CNBr) digests of the a polymer component of human fibrin has been isolated and characterized. The material has been used to generate monoclonal antibodies toward the goals of (1) producing fibrin-specific probes for use in the early detection of thrombosis and (2) generating monoclonal lines to single determinants in the COOH-terminal region of the Aα chain for use in structural studies of fibrinogen and fibrin.Biochemical and immunologic characterization data indicate the fragment is comprised, predominantly, of equimolar quantities of the CNBr peptides spanning residues #241-476 (CNBr 8) and #518-584 (CNBr 10) in the original Aα chain. The acceptor and donor units are crosslinked via an average of 2.8-3.2 ε-(γ-glutamyl) lysine bonds per mole of CNBr 8 + CNBr 10 producing heterogeneously sized fragments in the range of 80,000-200,000 daltons.Two types of monoclonal lines have been obtained. The first react with regions of primary structure and in one instance immunoreactivity could be localized to the Aa tryptic peptide #253-268. The second type appear to recognize conformational determinants as shown by one antibody that reacts well with the CNBr crosslinked fragment but poorly with its constituent peptides.

Download Full-text

Non-specific binding of mouse IgG1 to Heligmosomoides polygyrus: parasite homogenate can affinity purify mouse monoclonal antibodies

Parasitology ◽

10.1017/s0031182096008098 ◽

1997 ◽

Vol 114 (1) ◽

pp. 79-84 ◽

Cited By ~ 5

Author(s):

M. ROBINSON ◽

T. R. GUSTAD ◽

S. MEINHARDT

Keyword(s):

Monoclonal Antibodies ◽

Adult Worm ◽

Primary Infection ◽

Cyanogen Bromide ◽

Specific Binding ◽

Affinity Column ◽

Heligmosomoides Polygyrus ◽

Specific Proteins ◽

Mouse Lymphocytes

A characteristic feature of infections with the nematode parasite of mice Heligmosomoides polygyrus, is a marked IgG1 hypergammaglobulinaemia. A possible source for this immunoglobulin has recently been demonstrated, through evidence that H. polygyrus adult worm homogenate (AWH) can induce the in vitro production of non-specific IgG1 from mouse lymphocytes. To determine the interactions between this immunoglobulin and the parasite, the ability of IgG1 to bind to AWH of H. polygyrus was investigated. Protein (Western) blotting indicated that mouse monoclonal antibodies are able to bind non-specifically to selected parasite antigens. Furthermore, by binding H. polygyrus adult worm homogenate to cyanogen bromide (CNBr)-activated Sepharose CL-4B, an affinity column was prepared which could be used to efficiently purify mouse IgG1 monoclonal antibodies. These antibodies were eluted from the affinity column and still retained their original specificity. These results indicate that H. polygyrus not only induces the production of non-specific IgG1 by the host, it can also bind this immunoglobulin to its own specific proteins. Thus, it is possible that IgG1 produced during a primary infection with H. polygyrus may not entirely benefit the host.

Download Full-text

Ultrastructural analysis of unique glycoconjugates in the rat olfactory system

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100124859 ◽

1992 ◽

Vol 50 (1) ◽

pp. 890-891

Author(s):

James E. Crandall ◽

Linda C. Hassinger ◽

Gerald A. Schwarting

Keyword(s):

Central Nervous System ◽

Nervous System ◽

Monoclonal Antibodies ◽

Olfactory System ◽

Olfactory Bulbs ◽

Temporal And Spatial Patterns ◽

Carbohydrate Epitopes ◽

Olfactory Systems ◽

Temporal And Spatial ◽

Cell Surface Glycoconjugates

Cell surface glycoconjugates are considered to play important roles in cell-cell interactions in the developing central nervous system. We have previously described a group of monoclonal antibodies that recognize defined carbohydrate epitopes and reveal unique temporal and spatial patterns of immunoreactivity in the developing main and accessory olfactory systems in rats. Antibody CC2 reacts with complex α-galactosyl and α-fucosyl glycoproteins and glycolipids. Antibody CC1 reacts with terminal N-acetyl galactosamine residues of globoside-like glycolipids. Antibody 1B2 reacts with β-galactosyl glycolipids and glycoproteins. Our light microscopic data suggest that these antigens may be located on the surfaces of axons of the vomeronasal and olfactory nerves as well as on some of their target neurons in the main and accessory olfactory bulbs.

Download Full-text

Structural and Chemical Studies of Pathological Fibers in Human Neurofibrillary Degeneration

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010012031x ◽

1985 ◽

Vol 43 ◽

pp. 726-729

Author(s):

K.S. Kosik ◽

L.K. Duffy ◽

S. Bakalis ◽

C. Abraham ◽

D.J. Selkoe

Keyword(s):

Monoclonal Antibodies ◽

Alzheimer Disease ◽

Human Brain ◽

Neurofibrillary Tangles ◽

Morphological Analysis ◽

High Specificity ◽

Cytoskeletal Proteins ◽

Neuritic Plaque ◽

Protein Chemistry ◽

Chemical Studies

The major structural lesions of the human brain during aging and in Alzheimer disease (AD) are the neurofibrillary tangles (NFT) and the senile (neuritic) plaque. Although these fibrous alterations have been recognized by light microscopists for almost a century, detailed biochemical and morphological analysis of the lesions has been undertaken only recently. Because the intraneuronal deposits in the NFT and the plaque neurites and the extraneuronal amyloid cores of the plaques have a filamentous ultrastructure, the neuronal cytoskeleton has played a prominent role in most pathogenetic hypotheses.The approach of our laboratory toward elucidating the origin of plaques and tangles in AD has been two-fold: the use of analytical protein chemistry to purify and then characterize the pathological fibers comprising the tangles and plaques, and the use of certain monoclonal antibodies to neuronal cytoskeletal proteins that, despite high specificity, cross-react with NFT and thus implicate epitopes of these proteins as constituents of the tangles.

Download Full-text