SDS-PAGE of soluble (5 μl : A, C & F) and insoluble extracts (5 μl ; B, D & G) of the five best clones of scFv 225

<p><b>Copyright information:</b></p><p>Taken from "Directed evolution of single-chain Fv for cytoplasmic expression using the β-galactosidase complementation assay results in proteins highly susceptible to protease degradation and aggregation"</p><p>Microbial Cell Factories 2004;3():16-16.</p><p>Published online 17 Dec 2004</p><p>PMCID:PMC544847.</p><p>Copyright © 2004 Philibert and Martineau; licensee BioMed Central Ltd.</p>28S (A, B, C & D) and scFv HuLys11 (F & G) expressed in TG1 and TG1. Proteins were revealed either by Coomassie blue staining (A & B) or by western blot (C, D, F & G) using an HRP-coupled anti-polyHistidine monoclonal antibody (Sigma A7058) and detected with a commercial detection kit (Pierce Supersignal West Pico kit #14079). In panel E, the same soluble extracts than in C were tested for βgal activity

SDS-PAGE of soluble and insoluble extracts (5 μl) of the best clones isolated after four rounds of mutagenesis, cloned in the pET23NN plasmid

<p><b>Copyright information:</b></p><p>Taken from "Directed evolution of single-chain Fv for cytoplasmic expression using the β-galactosidase complementation assay results in proteins highly susceptible to protease degradation and aggregation"</p><p>Microbial Cell Factories 2004;3():16-16.</p><p>Published online 17 Dec 2004</p><p>PMCID:PMC544847.</p><p>Copyright © 2004 Philibert and Martineau; licensee BioMed Central Ltd.</p> Proteins were revealed either by Coomassie staining (top) or by western blot using the 9E10 anti-c-myc monoclonal antibody followed by an anti-mouse alkaline phosphatase-conjugated antibody and detected with the chromogenic substrate BCIP/NBT. A) HuLys11 scFv and mutants; B) 225.28S scFv and mutants

A focused antibody library for selecting scFvs expressed at high levels in the cytoplasm-0

<p><b>Copyright information:</b></p><p>Taken from "A focused antibody library for selecting scFvs expressed at high levels in the cytoplasm"</p><p>http://www.biomedcentral.com/1472-6750/7/81</p><p>BMC Biotechnology 2007;7():81-81.</p><p>Published online 22 Nov 2007</p><p>PMCID:PMC2241821.</p><p></p>rary (see additional file : Sequence of randomly picked clones)

Homology study showed that this amino acid was highly conserved through species for the c.1041C>A and c.1151C>T mutations.

<p>Homology study showed that this amino acid was highly conserved through species for the c.1041C>A and c.1151C>T mutations.</p

A focused antibody library for selecting scFvs expressed at high levels in the cytoplasm-2

<p><b>Copyright information:</b></p><p>Taken from "A focused antibody library for selecting scFvs expressed at high levels in the cytoplasm"</p><p>http://www.biomedcentral.com/1472-6750/7/81</p><p>BMC Biotechnology 2007;7():81-81.</p><p>Published online 22 Nov 2007</p><p>PMCID:PMC2241821.</p><p></p> positive and negative controls, respectively [5]. At 24 h post-transfection, cells were fixed and visualized under a fluorescent microscope with the fluorescein isothiocyanate filter set. The micrographs represent typical fields containing a similar number of cells in each case. Magnification: × 400

Clinical and hormonal data of patients with mutated <i>MAMLD1</i>.

<p>SD: standard deviation. ND: not determined. NA: not available. DHT: dihydrotestosterone. DHEA: dihydroepiandrsosterone. Parentheses indicate the standard deviation for height and weight and the normal range for hormone serum levels. Testes of 1–2 ml can be regarded as normal, as recently reported by Shibata et al. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032505#pone.0032505-Shibata1" target="_blank">[34]</a>.</p>*<p>It is notable that anti-mullerian hormone and inhibin were lowered in one case. <i>MAMLD1</i> is indeed reported to be expressed in Sertoli cells, as well <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032505#pone.0032505-Fukami1" target="_blank">[15]</a>.</p

A focused antibody library for selecting scFvs expressed at high levels in the cytoplasm-1

<p><b>Copyright information:</b></p><p>Taken from "A focused antibody library for selecting scFvs expressed at high levels in the cytoplasm"</p><p>http://www.biomedcentral.com/1472-6750/7/81</p><p>BMC Biotechnology 2007;7():81-81.</p><p>Published online 22 Nov 2007</p><p>PMCID:PMC2241821.</p><p></p>ed in under the control of the T7 promoter. Soluble extracts were prepared, separated by SDS-PAGE, and analyzed by Coomassie staining (a) or Western-blot (b) using 9E10 and an alkaline phosphatase conjugated anti-mouse IgG antibody (substrate BCIP/NBT). Each lane corresponded to 2 × 10cells. The arrow on the left indicates the position of the scFv

A focused antibody library for selecting scFvs expressed at high levels in the cytoplasm-4

<p><b>Copyright information:</b></p><p>Taken from "A focused antibody library for selecting scFvs expressed at high levels in the cytoplasm"</p><p>http://www.biomedcentral.com/1472-6750/7/81</p><p>BMC Biotechnology 2007;7():81-81.</p><p>Published online 22 Nov 2007</p><p>PMCID:PMC2241821.</p><p></p>es represent typical cells transfected with scFv13R4 and three representative anti-histones clones (2, 5 and 10). D, DAPI staining (blue) merged with the GFP signal

Incidence of exonic polymorphisms p.P359S and p.N662S, and relative haplotypes in normal controls and 46,XY DSD patients.

<p>Controls are combined with the published series (matched for ethnicity of patients and controls) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032505#pone.0032505-Kalfa1" target="_blank">[13]</a><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032505#pone.0032505-Chen1" target="_blank">[14]</a>. The χ-square test was performed. When combining all patients with the p.662G polymorphism whatever the p.359 allele, this p.662G was significantly more frequent in 46,XY DSD patients: 27.1% (n = 19) vs. 6.8% (n = 40), <i>p</i> = 0.0001.</p

Tertiary structure prediction of the wildtype protein (left column) and with the mutants.

<p>3D structure was predicted at Protein Homology/analogY Recognition Engine (PhyreEngine) from the Structural Bioinformatics Group, Imperial College, London, at <a href="http:www.sbg.bio.ic.ac.uk/phyre~/" target="_blank">http:www.sbg.bio.ic.ac.uk/phyre~/</a>. The plain arrows show the changes in the shape of the protein between the wildtype and p.P384L.</p