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Results: 5

1.
Figure 3

Figure 3. From: Generation of monospecific antibodies based on affinity capture of polyclonal antibodies.

Generation of monospecific antibodies towards the human transcription factor SATB2. Each panel shows the epitopes, the molecular identification in Western blot (Lane 1: Caco-2 cell lysate), staining of colon cancer tissue, and subcellular localization in CACO-2 cells for the polyclonal sera purified from tag specific antibodies (pAb) targeting SATB2 and five monospecific and one conformational antibody purified from polyclonal sera, respectively.

Barbara Hjelm, et al. Protein Sci. 2011 November;20(11):1824-1835.
2.
Figure 1

Figure 1. From: Generation of monospecific antibodies based on affinity capture of polyclonal antibodies.

The principle for generation of monospecific antibodies. The polyclonal serum is assayed with a peptide array spanning the antigen sequence to identify the epitopes of the antibodies (A). The epitope corresponding peptides as well as the recombinant antigen are coupled to columns which are mounted in a serial manner after a protein tag specific column onto which the polyclonal serum is applied (B). Bound antibodies are eluted in parallel to create monospecific antibodies which are evaluated in different paired antibody applications (C).

Barbara Hjelm, et al. Protein Sci. 2011 November;20(11):1824-1835.
3.
Figure 4

Figure 4. From: Generation of monospecific antibodies based on affinity capture of polyclonal antibodies.

Generation of monospecific antibodies towards the human enzyme ANLN. Each panel shows the epitopes, the molecular identification in Western blot (Lane 1: RT-4 cell lysate Lane 2: U-251 MG cell lysate), staining of breast cancer tissue, and subcellular localization in U-251 MG cells for the polyclonal purified from tag specific antibodies (pAb), four monospecific and one conformational antibody targeting ANLN respectively. To the right, staining of ANLN siRNA treated U-251 MG cells, microtubuli (red), nucleus (blue), and respective antibody in green.

Barbara Hjelm, et al. Protein Sci. 2011 November;20(11):1824-1835.
4.
Figure 2

Figure 2. From: Generation of monospecific antibodies based on affinity capture of polyclonal antibodies.

Generation of monospecific antibodies towards the human RNA-binding protein RBM3. Each panel shows the epitopes, the molecular identification in Western blot (Lane 1: RT-4 cell lysate Lane 2: U-251 MG cell lysate), staining of breast cancer tissue, and subcellular localization in U-251 MG cells (images are shown in a rainbow RGB look up table (LUT) where red indicates strong signal and blue low signal) for the polyclonal serum purified from tag specific antibodies (pAb) targeting RBM3, five monospecific and one conformational antibody purified from the polyclonal sera, respectively. The peptide selected for purification are indicated in the epitope mapping barplots in different colors, where the x-axis represents the peptide ID and the y-axis indicate the mean fluorescence intensity. A small plus indicate the detection of a band of correct molecular mass, the right staining or localization according to literature while a minus indicate aberrant or negative result.

Barbara Hjelm, et al. Protein Sci. 2011 November;20(11):1824-1835.
5.
Figure 5

Figure 5. From: Generation of monospecific antibodies based on affinity capture of polyclonal antibodies.

Generation of monospecific antibodies towards the human enzyme CNDP1. A: In the left panel, epitopes and molecular identification in Western blot (Lane 1: Purified recombinant protein CNDP1) is shown for the polyclonal sera purified from tag specific antibodies (pAb) and five monospecific antibodies purified from the sera. To the right, titration curves resulting from the detection of CNDP1 spiked into plasma are shown. Sandwich pairs built on monospecific antibodies were compared using a 6-plex bead array, and two detection antibody candidates are presented in combination with all six antibodies as capture reagents. Rating a pair as functional was based on a relative detection limit for CNDP1 <10 ng/mL and is indicated with a plus. B: Crystal structure of CNDP1 (http://www.pdb.org/pdb/explore/explore.do?pdbId=3DLJ) with peptide 1 (cyan) and peptide 15 (green) indicated. Below a linear view of the amino acid sequence of CNDP1 with the antigen used for generation of polyclonal antibody marked in grey.

Barbara Hjelm, et al. Protein Sci. 2011 November;20(11):1824-1835.

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