Generation of human scFvs antibodies recognizing a prion protein epitope expressed on the surface of human lymphoblastoid cells-0

<p><b>Copyright information:</b></p><p>Taken from "Generation of human scFvs antibodies recognizing a prion protein epitope expressed on the surface of human lymphoblastoid cells"</p><p>http://www.biomedcentral.com/1472-6750/7/38</p><p>BMC Biotechnology 2007;7():38-38.</p><p>Published online 2 Jul 2007</p><p>PMCID:PMC1933425.</p><p></p>y populations with CCRF-CEM cells are shown after each round of biopanning selection. In the inserted boxes, the ELISA reactivity of the ETH-2 library (A') and selected phage antibody populations (B', C', D') with the rHaPrP are shown in parallel with the irrelevant phage antibody anti glucose oxidase and anti tetanus toxoid (A", B", C" and D"). Experiments were repeated at least twice and mean ± SD from representative experiments (triplicate samples) is shown. The cut-off value separating positive from negative sample was calculated as 3 standard deviation above the mean of the value obtained from irrelevant phage antibodies (0,085 OD). In the lower part of the figure, the nucleotide composition and the corresponding amino acid sequences in the CDR3 regions of the selected scFv antibodies MA3.B4 and MA3.G3 are shown. A schematic representation of the scFv antibody gene as M13 pIII fusion protein is also illustrated

Generation of human scFvs antibodies recognizing a prion protein epitope expressed on the surface of human lymphoblastoid cells-1

<p><b>Copyright information:</b></p><p>Taken from "Generation of human scFvs antibodies recognizing a prion protein epitope expressed on the surface of human lymphoblastoid cells"</p><p>http://www.biomedcentral.com/1472-6750/7/38</p><p>BMC Biotechnology 2007;7():38-38.</p><p>Published online 2 Jul 2007</p><p>PMCID:PMC1933425.</p><p></p>tion. As controls the irrelevant phage antibody anti glucose oxidase and anti tetanus toxoid are also shown. Experiments were repeated at least twice and mean ± SD from representative experiments (triplicate samples) is shown. The cut-off value separating positive from negative sample was calculated as 3 standard deviation above the mean of the value obtained from irrelevant phage antibodies (0,086 OD for rHaPrp, 0,052 OD for brain homogenate). In panel B, the monoclonal phage antibodies and the rodent mAb 3F4 were tested in flow-cytometry on human HL-60 cells. The surface PrPc expression was down-regulated by ATRA inducing granulocytic differentiation. HL-60 cells were cultured in the presence of 1 μM ATRA for 1 and 3 days, then stained for flow cytometry analysis with scFvs MA3.B4, MA3.G3 and mAb 3F4. Basal level of PrPc staining in undifferentiated cells (solid line) is shown relative to an irrelevant phage antibody, specific for glucose oxidase, or an IgG2b isotype control (in gray). The progressive decrease of PrPc staining during differentiation is shown in black

Generation of human scFvs antibodies recognizing a prion protein epitope expressed on the surface of human lymphoblastoid cells-3

<p><b>Copyright information:</b></p><p>Taken from "Generation of human scFvs antibodies recognizing a prion protein epitope expressed on the surface of human lymphoblastoid cells"</p><p>http://www.biomedcentral.com/1472-6750/7/38</p><p>BMC Biotechnology 2007;7():38-38.</p><p>Published online 2 Jul 2007</p><p>PMCID:PMC1933425.</p><p></p> at two different magnification, in C and D the reactivity of CCRF-CEM cells with two distinct irrelevant phage antibodies (anti glucose oxidase and anti tetanus toxoid, respectively) are shown

Generation of human scFvs antibodies recognizing a prion protein epitope expressed on the surface of human lymphoblastoid cells-4

<p><b>Copyright information:</b></p><p>Taken from "Generation of human scFvs antibodies recognizing a prion protein epitope expressed on the surface of human lymphoblastoid cells"</p><p>http://www.biomedcentral.com/1472-6750/7/38</p><p>BMC Biotechnology 2007;7():38-38.</p><p>Published online 2 Jul 2007</p><p>PMCID:PMC1933425.</p><p></p>y populations with CCRF-CEM cells are shown after each round of biopanning selection. In the inserted boxes, the ELISA reactivity of the ETH-2 library (A') and selected phage antibody populations (B', C', D') with the rHaPrP are shown in parallel with the irrelevant phage antibody anti glucose oxidase and anti tetanus toxoid (A", B", C" and D"). Experiments were repeated at least twice and mean ± SD from representative experiments (triplicate samples) is shown. The cut-off value separating positive from negative sample was calculated as 3 standard deviation above the mean of the value obtained from irrelevant phage antibodies (0,085 OD). In the lower part of the figure, the nucleotide composition and the corresponding amino acid sequences in the CDR3 regions of the selected scFv antibodies MA3.B4 and MA3.G3 are shown. A schematic representation of the scFv antibody gene as M13 pIII fusion protein is also illustrated

Generation of human scFvs antibodies recognizing a prion protein epitope expressed on the surface of human lymphoblastoid cells-2

<p><b>Copyright information:</b></p><p>Taken from "Generation of human scFvs antibodies recognizing a prion protein epitope expressed on the surface of human lymphoblastoid cells"</p><p>http://www.biomedcentral.com/1472-6750/7/38</p><p>BMC Biotechnology 2007;7():38-38.</p><p>Published online 2 Jul 2007</p><p>PMCID:PMC1933425.</p><p></p>mphoblastoid, myeloid and neuronal origin are shown in respect to an irrelevant phage antibody, specific for glucose oxidase, or to an IgG2b isotype control (solid line)

Selection, affinity maturation, and characterization of a human scFv antibody against CEA protein-0

<p><b>Copyright information:</b></p><p>Taken from "Selection, affinity maturation, and characterization of a human scFv antibody against CEA protein"</p><p>BMC Cancer 2006;6():41-41.</p><p>Published online 24 Feb 2006</p><p>PMCID:PMC1402309.</p><p>Copyright © 2006 Pavoni et al; licensee BioMed Central Ltd.</p>uration library was constructed by error-prone PCR method

Selection, affinity maturation, and characterization of a human scFv antibody against CEA protein-8

<p><b>Copyright information:</b></p><p>Taken from "Selection, affinity maturation, and characterization of a human scFv antibody against CEA protein"</p><p>BMC Cancer 2006;6():41-41.</p><p>Published online 24 Feb 2006</p><p>PMCID:PMC1402309.</p><p>Copyright © 2006 Pavoni et al; licensee BioMed Central Ltd.</p

Selection, affinity maturation, and characterization of a human scFv antibody against CEA protein-6

<p><b>Copyright information:</b></p><p>Taken from "Selection, affinity maturation, and characterization of a human scFv antibody against CEA protein"</p><p>BMC Cancer 2006;6():41-41.</p><p>Published online 24 Feb 2006</p><p>PMCID:PMC1402309.</p><p>Copyright © 2006 Pavoni et al; licensee BioMed Central Ltd.</p>of the negative control scFv directed to glucose oxidase

Selection, affinity maturation, and characterization of a human scFv antibody against CEA protein-1

<p><b>Copyright information:</b></p><p>Taken from "Selection, affinity maturation, and characterization of a human scFv antibody against CEA protein"</p><p>BMC Cancer 2006;6():41-41.</p><p>Published online 24 Feb 2006</p><p>PMCID:PMC1402309.</p><p>Copyright © 2006 Pavoni et al; licensee BioMed Central Ltd.</p>bated with irrelevant anti-glucose oxidase (a protein not present in mammals) scFv antibody (anti-G.O., right slide). B. Biochemical patterns of the MA39 on purified CEA (lane 1) and LoVo cellular extract (lane 2). Lanes 3 and 4 show the reactivity of identical proteins with irrelevant anti-G.O. scFv. C. Staining of cryosections from human colon carcinoma xenograft in mouse. Commercially available anti-CEA antibody and irrelevant anti-G.O. antibodies were used as positive and negative controls, respectively

Selection, affinity maturation, and characterization of a human scFv antibody against CEA protein-4

<p><b>Copyright information:</b></p><p>Taken from "Selection, affinity maturation, and characterization of a human scFv antibody against CEA protein"</p><p>BMC Cancer 2006;6():41-41.</p><p>Published online 24 Feb 2006</p><p>PMCID:PMC1402309.</p><p>Copyright © 2006 Pavoni et al; licensee BioMed Central Ltd.</p>s. Significant fluorescence shift indicates difference in affinity of two single-chain antibodies. B. Western blot with serial dilutions of CEA protein was stained by using original MA39 and maturated E8 scFv antibodies. The membranes were developed with secondary anti-FLAG AP-conjugated antibody