Quantification of the secretory glycoproteins of the subcommissural organ by a sensitive sandwich ELISA with a polyclonal antibody and a set of monoclonal antibodies against the bovine Reissner's fiber

1998 ◽  
Vol 294 (3) ◽  
pp. 407-413 ◽  
Author(s):  
G. Estivill-Torr�s ◽  
M. Cifuentes ◽  
E. Miranda ◽  
F. J. Berm�dez-Silva ◽  
P. Fern�ndez-Llebrez ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Polyclonal Antibody ◽  
Subcommissural Organ ◽  
Sandwich Elisa ◽  
Reissner's Fiber ◽  
Reissner’S Fiber ◽  
Secretory Glycoproteins

Isograft and xenograft of the subcommissural organ into the lateral ventricle of the rat and the formation of Reissner's fiber

1999 ◽  
Vol 296 (3) ◽  
pp. 457-469 ◽  
Author(s):  
S. Rodríguez ◽  
E. H. Navarrete ◽  
K. Vio ◽  
C. González ◽  
K. Schöbitz ◽  
...  
Keyword(s):  
Lateral Ventricle ◽  
Subcommissural Organ ◽  
Reissner's Fiber ◽  
Reissner’S Fiber

Reissner’s fiber formation depends on developmentally regulated factors extrinsic to the subcommissural organ

2005 ◽  
Vol 321 (3) ◽  
pp. 429-441 ◽  
Author(s):  
C. Hoyo-Becerra ◽  
M. D. López-Avalos ◽  
M. Alcaide-Gavilán ◽  
M. C. Gómez-Roldán ◽  
J. Pérez ◽  
...  
Keyword(s):  
Subcommissural Organ ◽  
Fiber Formation ◽  
Developmentally Regulated ◽  
Reissner's Fiber ◽  
Reissner’S Fiber

Identification of Reissner's fiber-like glycoproteins in two species of freshwater planarians (Tricladida), by use of specific polyclonal and monoclonal antibodies

2000 ◽  
Vol 300 (3) ◽  
pp. 427-434
Author(s):  
P. M. Arrabal ◽  
G. Estivill-Torrús ◽  
E. Miranda ◽  
J. Pérez ◽  
P. Fernández-Llebrez
Keyword(s):  
Monoclonal Antibodies ◽  
Reissner's Fiber ◽  
Reissner’S Fiber

Hydrocephalus induced by immunological blockage of the subcommissural organ-Reissner's fiber (RF) complex by maternal transfer of anti-RF antibodies

2000 ◽  
Vol 135 (1) ◽  
pp. 41-52 ◽  
Author(s):  
K. Vio ◽  
S. Rodríguez ◽  
E.H. Navarrete ◽  
J.M. Pérez-Fígares ◽  
A.J. Jiménez ◽  
...  
Keyword(s):  
Subcommissural Organ ◽  
Maternal Transfer ◽  
Reissner's Fiber ◽  
Reissner’S Fiber

Development of a Sandwich ELISA to Detect Hepcidin in Human Serum.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 2000-2000 ◽  
Author(s):  
Tara L Arvedson ◽  
George Doellgast ◽  
Hossein Salimi-Moosavi ◽  
Chadwick King ◽  
Ian Foltz ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Amino Acid ◽  
Polyclonal Antibody ◽  
Limit Of Detection ◽  
Sandwich Elisa ◽  
Alkaline Treatment ◽  
Detection Range ◽  
Antibody Preparation ◽  
Amino Acid Peptide ◽  
Class 1

Abstract Abstract 2000 Poster Board I-1022 Hepcidin is a 25 amino acid peptide that is the central mediator of iron metabolism. Iron excess, deficiency and maldistribution have been implicated in the etiology of many diseases including atherosclerosis, diabetes, neurodegeneration and the anemia of inflammation. Determination of hepcidin levels may be useful in diagnosis and treatment decisions for some or all of these diseases. Serum hepcidin measurement has so far been limited to a prohepcidin (60 amino acid hepcidin precursor) ELISA, mass spectrometry (MS)-based assays or competition ELISAs using polyclonal anti-hepcidin antibodies. The current work describes the generation of a sandwich ELISA using monoclonal antibodies to detect human hepcidin (hHepc) and optimization of assay conditions to resolve inconsistencies between MS- and ELISA-based detection. The ability of two anti-hHepc antibodies to sandwich (bind simultaneously) with hHepc was demonstrated using a rabbit polyclonal antibody preparation from hHepc-immunized animals. The same polyclonal antibody preparation was used for both hHepc capture and detection. The limit of detection achieved with this assay was O.D.450<1, suggesting that only a small proportion of the total antibodies could bind concurrently. To improve hHepc detection, a panel of monoclonal antibodies was screened for the ability to sandwich. Antibody epitope characterization studies using purified antibodies and >1000 hybridoma supernatants identified three classes of antibodies: classes 1 and 2 each recognized epitopes found in both full length mature hHepc (hHepc 25) and a shorter version (hHepc 20); class 3 bound a different epitope and demonstrated an increased affinity for hHepc 25 over hHepc 20. The majority of antibodies characterized were in class 1 while antibodies in classes 2 and 3 were rare (∼1% of antibody panel) highlighting the difficulty in achieving a sandwiching event. Antibodies 19D12 (class 1) and 23F11 (class 2) were identified as the optimal sandwich pair with a detection range of approximately 0.2-1000 ng/ml using synthetic hHepc. Initial comparisons of data generated using the sandwich ELISA and a fully-quantitative MS-based assay demonstrated a lack of consistent agreement. This issue was somewhat addressed by introduction of an alkaline treatment step to dissociate any protein/hHepc complexes in serum. Subsequent comparison of the two assays using sera from several different patient populations (anemia of cancer, chemotherapy-induced anemia, kidney disease) as well as healthy donors demonstrated good correlation (R2 range = 0.83-0.92; n=237). This sandwich ELISA may represent a tool for aligning the MS and ELISA-generated results in a format that has the potential to be high throughput and widely available. Disclosures: Arvedson: Amgen: Employment. Doellgast:Amgen: Employment. Salimi-Moosavi:Amgen: Employment. King:Amgen: Employment. Foltz:Amgen: Employment. Chen:Amgen: Employment. Li:Amgen: Employment. Sasu:Amgen: Employment.

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