Analysis of surface properties of fixed and live cells using derivatized agarose beads

Navarro, Vanessa M.; Walker, Sherri L.; Badali, Oliver; Abundis, Maria I.; Ngo, Lylla; Weerasinghe, Gayani; Barajas, Marcela; Zem, Gregory; Oppenheimer, Steven B.

doi:10.1078/0065-1281-00617

Cited by 18 publications

(21 citation statements)

References 11 publications

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“…Binding was confirmed by agitating the suspension, observing if the cells were bound to the lectin beads or were simply touching them. All assays were conducted in distilled water as this has been shown to be a reliable medium for such studies (Roque et al, 1996;Navarro et al, 2002;Khurrum et al, 2002;Welty et al, 2006).…”

Section: Lectin Bead Binding Assaymentioning

confidence: 99%

“…We have tested the ability of cells to bind to lectin-derivatized agarose beads using yeast (Saccharomyces cerevisiae) cells (Salbilla et al, 1999;Navarro et al, 2002) or sea urchin embryos (Strongylocentrotus purpuratus) (Latham et al, 1995a;1995b; and recently we have examined the cell surface properties of human colon cancer (CCL-220, CCL-255), human lung cancer (HTB-171) and human non-cancer colon (CRL-1459) cells (Khurrum et al, 2002;Heinrich et al, 2005;Welty et al, 2006). Few past studies have examined all three of the parameters that we examine in this model system: cell binding of the agent, toxicity of the agent and testing both cancer and non-cancer cells of the same tissue type (Valentiner et al, 2003;Heinrich et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

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Lectin binding and effects in culture on human cancer and non-cancer cell lines: Examination of issues of interest in drug design strategies

2007

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SummaryBy using a non-cancer and a cancer cell line originally from the same tissue (colon), coupled with testing lectins for cell binding and for their effects on these cell lines in culture, this study decribes a simple multiparameter approach that has revealed some interesting results that could be useful in drug development strategies.Two human cell lines, CCL-220/Colo320DM (human colon cancer cells, tumorigenic in nude mice) and CRL-1459/CCD-18Co (non-malignant human colon cells) were tested for their ability to bind to agarose microbeads derivatized with two lectins, peanut agglutinin (Arachis hypogaea agglutinin, PNA) and Dolichos biflorus agglutinin (DBA), and the effects of these lectins were assessed in culture using the MTT assay.Both cell lines bound to DBA-derivatized microbeads, and binding was inhibited by N-acetyl-Dgalactosamine, but not by L-fucose. Neither cell line bound to PNA-derivatized microbeads. Despite the lack of lectin binding using the rapid microbead method, PNA was mitogenic in culture at some time points and its mitogenic effect displayed a reverse-dose response. This was also seen with effects of DBA on cells in culture.

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Section: Lectin Bead Binding Assaymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Lectin binding and effects in culture on human cancer and non-cancer cell lines: Examination of issues of interest in drug design strategies

2007

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“…Many past studies have used derivatized beads to examine cell surface properties (Goldstein et al 1976;Oketa et al 1979;Iweg et al 1980;Takahashi and Tavassoli 1981;Nenci 1983;Pincus 1984;Johnson and Silver 1989;Matsuoka and Tavassoli 1989;Hayes et al 1990; Kijimoto-Ochiai and Uede 1995), but we are unaware of any studies, other than ours, that use so many different beads to rapidly assay cell surface properties (Khurrum et al 2002;Navarro et al 2002;Salbilla et al 1999;Roque et al 1996;Latham and Oppenheimer, 1999;Latham et al 1995;1995a;Heinrich et al 2005). Previous studies from our laboratory have involved many unconventional approaches.…”

Section: Introductionmentioning

confidence: 87%

“…Over the past decade, this laboratory has developed and used a novel histochemical assay to examine cell surface properties (Heinrich, et al 2005;Khurrum, et al 2002;Navarro et al 2002;Salbilla et al 1999;Latham, et al 1995;1995a;Latham and Oppenheimer, 1999). This *To whom correspondence should be addressed.…”

Section: Introductionmentioning

confidence: 99%

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Analysis of unconventional approaches for the rapid detection of surface lectin binding ligands on human cell lines

et al. 2006

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SummaryThis laboratory has developed and used a novel histochemical assay using derivatized agarose beads for over a decade to examine the surface properties of various cell types. Most recently we have used this assay to examine lectin binding ligands on two human cell types, CCL-220, a colon cancer cell line, and CRL-1459, a non-cancer colon cell line. We found that CCL-220 cells bound specific lectins better than CRL-1459, and this information was used to test for possible differential toxicity of these lectins in culture, as a possible approach in the design of more specific anti-cancer drugs. Although we have examined the validity of the bead-binding assay in sea urchin cell systems, we have not validated this technique for mammalian cells. Here the binding results of the bead assay are compared with conventional fluorescence assays, using lectins from 3 species (Triticum vulgaris, Phaseolus vulgaris, and Lens culinaris) on the 2 colon cell lines. These lectins were chosen because they seemed to interact with the two cell lines differently.Binding results obtained using both assays were compared for frozen, thawed and fixed; cultured and fixed; and live cells. Both qualitative and quantitative fluorescence results generally correlated with those using the bead assay. Similar results were also obtained with all of the 3 different cell preparation protocols. The fluorescence assay was able to detect lower lectin binding ligand levels than the bead assay, while the bead assay, because it can so rapidly detect cells with large numbers of lectin binding ligands, is ideal for initial screening studies that seek to identify cells that are rich in surface binders for specific molecules. The direct use of frozen, thawed, and fixed cells allows rapid mass screening for surface molecules, without the requirement for costly and time consuming cell culture.

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Metabolically Stable Cellular Adhesion to Inert Surfaces

Meldal

Diness

et al. 2011

ChemBioChem

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The structure of D-amino acid hexapeptides that promote cellular adhesion was determined by screening D-amino acid hexapeptide libraries synthesized on otherwise inert beaded PEGA resin. These new adhesion molecules provide a completely stable cellular environment and facilitate the maintenance of a monolayer of cells on beads for extended periods. The presence of the peptides promotes spreading of the cells on the bead surface. Not surprisingly, the molecules contained a significant number of arginines and/or lysines. However, the exact structure of each peptide is quite important for the degree of adhesion observed, and a motif with three or four basic amino acids spaced within amino acids of intermediate polarity clearly prevailed, for example, k-l/r-h-r-i/v-r-a; this maintains a polar/hydrophobic balance.

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Analysis of surface properties of fixed and live cells using derivatized agarose beads

Cited by 18 publications

References 11 publications

Lectin binding and effects in culture on human cancer and non-cancer cell lines: Examination of issues of interest in drug design strategies

Lectin binding and effects in culture on human cancer and non-cancer cell lines: Examination of issues of interest in drug design strategies

Analysis of unconventional approaches for the rapid detection of surface lectin binding ligands on human cell lines

Metabolically Stable Cellular Adhesion to Inert Surfaces

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