Comparative studies of iodo-bead and chloramine-T methods for the radioiodination of human alpha-fetoprotein

Lee, David S.C.; Griffiths, B. W.

doi:10.1016/0022-1759(84)90379-x

Cited by 39 publications

(10 citation statements)

References 11 publications

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“…F(ab')2 fragments of MoAb SW73.2 were iodinated with '"''iodine (Amersham Int,. Aylesbury, UK) using the Iodobeads iodination protocol (Pierce, Rockford, IL) [12]. The level of expression of MHC class II molecules was measured by saturation binding using '"^I-labelled SW73,2 F(ab'); as previously described [13,14], except that cells were labelled on ice for 30 min before unbound F(ab')> was separated from the cells by washing with aliquots of PBS, The results obtained from assays of the cloned bovine B cell lytnphoma line BL20 were identical to those previously described [15].…”

Section: Quantification Of Mhc Class II Expressionmentioning

confidence: 99%

T cell activation by Theileria annulata-infected macrophages correlates with cytokine production

Brown

Campbell

Russell

et al. 1995

Clinical and Experimental Immunology

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SUMMARYA major feature of the pathology induced by Theileria annulata is acute lymphocytic proliferation, and this study investigates the mechanisms underlying the intrinsic ability of T. anniilata-infecXed monocyles to induce naive autoiogous T cells to proliferate. Different 7". aiinulata-'mkcted clones expressed different but constant levels of MHC class II, varying from < 10 x 10' to 1-5 x 10niolecules/celK as measured by saturation binding. However, no correlation was found between the level of MHC class IT expression and levels of induced T ceil proliferation. Theileria animtatainfected cell lines and clones were assayed for cytokine mRNA expression by reverse transcriplionpolymerase chain reaction (RT-PCR). The infected cells assayed produced mRNA specific for IL-la. \L-\fi. lL-6, IL-10 and tumour necrosis factor-alpha (TNF-a). but not IL-2 or lL-4. One clone (clone G)did not produce mRNA for TNF-f>. The degree of Tcell proliferation induced by infected cells was directly correlated with the amount of mRNA produced for the T cell stimulatory cytokines IL-la and IL-6, as assessed by a semiquantitative technique. In contrast, cells infected with the related parasite T. parva produced mRNA for IL-la, IL-2, IL-4, IL-IO and interferon-gamma (IFN-7). Since T. /5(^/rv«-infected cells also induce naive autologous T cell proliferation, it seems likely that the production oflL-Ifi by cells infected with either parasite is a major signal for the induction of non-specific T cell proliferation.

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Section: Quantification Of Mhc Class II Expressionmentioning

confidence: 99%

T cell activation by Theileria annulata-infected macrophages correlates with cytokine production

Brown

Campbell

Russell

et al. 1995

Clinical and Experimental Immunology

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“…SHaPrPC was purified from the brains of uninoculated adult SHa (5) and radioiodinated with 1251 (1 mCi per 100 ,ug of PrPC; Amersham) using lodo-Beads (Pierce) (29).…”

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confidence: 99%

Prion protein (PrP) synthetic peptides induce cellular PrP to acquire properties of the scrapie isoform.

Kaneko

Peretz

Pan

et al. 1995

Proc. Natl. Acad. Sci. U.S.A.

103

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Conversion of the cellular isoform of prion protein (PrPC) into the scrapie isoform (PrPSc) involves an increase in the beta-sheet content, diminished solubility, and resistance to proteolytic digestion. Transgenetic studies argue that PrPC and PrPSc form a complex during PrPSc formation; thus, synthetic PrP peptides, which mimic the conformational pluralism of PrP, were mixed with PrPC to determine whether its properties were altered. Peptides encompassing two alpha-helical domains of PrP when mixed with PrPC produced a complex that displayed many properties of PrPSc. The PrPC-peptide complex formed fibrous aggregates and up to 65% of complexed PrPC sedimented at 100,000 x g for 1 h, whereas PrPC alone did not. These complexes were resistant to proteolytic digestion and displayed a high beta-sheet content. Unexpectedly, the peptide in a beta-sheet conformation did not form the complex, whereas the random coil did. Addition of 2% Sarkosyl disrupted the complex and rendered PrPC sensitive to protease digestion. While the pathogenic A117V mutation increased the efficacy of complex formation, anti-PrP monoclonal antibody prevented interaction between PrPC and peptides. Our findings in concert with transgenetic investigations argue that PrPC interacts with PrPSc through a domain that contains the first two putative alpha-helices. Whether PrPC-peptide complexes possess prion infectivity as determined by bioassays remains to be established.

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“…More recently, iodogen (1,3,4,6-tetrachloro-3a,6a-diphenylglycouril), which performs a mild oxidation of iodide, has been introduced (Fraker and Speck, 1978). In a manner similar to lactoperoxidase, iodogen produces a mildly iodinated molecule that exhibits minimal damage and high immunoreactivity at the expense of very high specific activities (Lee and Griffiths, 1984). For molecules that have no tyrosines or have tyrosines in binding regions of the molecule, the Bolton-Hunter reagent (an iodinated /?-hydroxyphenylpropionic acid, N-hydroxysuccinimide ester) has been used successfully.…”

Section: A Isotopic Labelsmentioning

confidence: 99%

“…An overview of separation methods most commonly employed in heterogeneous immunoassays is presented in Table IV. A. Liquid-Phase Methods Separation of free and bound labeled ligand or receptor was accomplished in many early immunoassays by differential adsorption of the labeled reagent to a solid material. Examples of this separation method include the use of cellulose powder (Zaharko and Beck, 1968), dextran-coated charcoal (Odell, 1980), ion- Zaharko and Beck, 1968Odell, 1980Lazarus and Young, 1966Franchimont et al, 1969Rosselin et al, 1966Makulu et al, 1969Chard, 1980Thomas and Ferin, 1968Chard, 1980Coffey et al, 1980Midgley and Hepburn, 1980Seibel et al, 1981Chalkley and Renshaw, 1980Freytag et al, 1984Light et al, 1977Wide and Porath, 1966Pourfarzaneh et al, 1982Kerschensteiner, 1984Masson et al, 1983Kung and Martin, 1984mW'dnetal, 1985Djurup, 1983 Ziola et al, \911 Odstrchel, 1980Borsos and Langone, 1983Lindmark, 1982Litchfield et al, 1984Zick et al, 1980Miller era/., 1984Pick and Wagner, 1980Hamaguchi et al, 1976Giaever, 1973Clark and Engvall, 1980Pappas et al, 1983Walter et al, 1983Catt and Tregear, 1967Sedlacek et al, 1983 exchange resins …”

Section: Separation Techniquesmentioning

confidence: 99%

Production of Immunoassay Reagents

Hamilton¹

1987

Immunoassay

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Comparative studies of iodo-bead and chloramine-T methods for the radioiodination of human alpha-fetoprotein

Cited by 39 publications

References 11 publications

T cell activation by Theileria annulata-infected macrophages correlates with cytokine production

T cell activation by Theileria annulata-infected macrophages correlates with cytokine production

Prion protein (PrP) synthetic peptides induce cellular PrP to acquire properties of the scrapie isoform.

Production of Immunoassay Reagents

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