KAPTIV A product line designed for the affinity purification of monoclonal or polyclonal antibodies from different sources. Ligands are synthetic peptidomimetic molecules, stable to repeated use and ideal for analytical and preparative uses. Selective matrices are available for IgG, IgM, IgA, IgE and IgY purification KAPTIV-AETM is available directly from TECNOGEN s.c.p.a. in different formats product code description TG9001 TG9002 TG9003 TG9004 KAPTIV-AETM KAPTIV-AETM KAPTIV-AETM KAPTIV-AETM KAPTIV-AE TM Af f inity matrix f o r I g A and IgE p ur i f i c a t i o n (5 ml) (10 ml) (25 ml) (100 ml) KAPTIV-AETM is available also in bulk amounts for preparative applications. Please contact our sales department for quotations and additional products of the KAPTIV line. KAPTIV-MTM Affinity matrix for IgM purificaiton KAPTIV-M product code description TG8001 TG8002 TG8003 TG8004 KAPTIV-MTM KAPTIV-MTM KAPTIV-MTM KAPTIV-MTM ™ (5 ml) (10 ml) (25 ml) (100 ml) to order Orders may be placed by phone, fax or E-mail directly to: TECNOGEN s.c.p.a. Parco Scientifico, Località La Fagianeria 81015 Piana di Monte Verna (CE) - ITALY Phone: +39-0823-612-208/228, Fax: +39-0823-612-222/230 E-mail: [email protected] Home page: http://www.tecnogen.it Capture of IgA and IgE made easy TECNOGEN s.c.p.a. KAPTIV-AETM Ready to use matrix for IgA and IgE purification Purification of IgA from serum samples, such as cell culture supernatants, ascites, or sera. Sample loading is achieved at room temperature with physiological buffers, and dissociation of bound immunoglobulins is achieved by a buffer change to 0.1 M acetic acid, 0.2 M glycine pH 2.2, or 0.1 M sodium bicarbonate pH 9.0, with full retention of immunological properties. Matrix capacity is at least 10 mg/IgA or IgE per ml of resin. KAPTIV-AE™ can be used in the batch mode or easily packed in columns for analytical as well as preparative applications, for a cost effective isolation of IgA or IgE with high purity degree. KAPTIV-AE™ is particularly useful for the down stream processing of monoclonal antibodies for therapy, given the high capacity and ligand stability. Recovery of IgA immunoreactivity from serum after affinity purification on KAPTIV-AE TM monitored by ELISA. A crude sample of mouse serum (0.2 ml) has been loaded on a KAPTIVAE™ column (2 ml) equilibrated at a flow rate of 1.0 ml/min with phosphate saline buffer (PBS) at room temperature. Adsorbed IgA was eluted by a buffer change to 0.1 M acetic acid and immediately neutralized. The majority of IgA immunoreactivity is found in the column adsorbed fraction (peak 2), while negligible activity is found in the column flow-through. Purification of monoclonal IgA from cell culture supernatants Immunoreactivity recovery (%) KAPTIV-AE™ is a new matrix optimized for the affinity purification of monoclonal or polyclonal IgA and IgE from crude peak 2 100 80 60 40 20 peak 1 0 peak 1 peak 2 190 First lane crude hybridoma supernatant. Second lane Unbound material from the KAPTIV-AE™ affinity column (peak 1) Third lane KAPTIV-AE™ affinity purified IgA (peak 2) ABS 280 nm - 0.002 A.U.F.S. Recovery of IgE immunoreactivity after affinity purification on KAPTIV-AETM. IgA crude IgA peak 2 0 Background: Structure of TG19318*, affinity ligand for IgA and IgE purification [*patents pending] 50 100 150 Elution time [min] 200 2 ak pe ak 190 First lane Crude ascitic fluid Second lane Column unretained material, only traces of IgE are visible (peak 1) Third lane KAPTIV-AE™ affinity purified IgE (peak 2) 66 serum albumin Gel permeation analysis of mouse monoclonal IgA affinity purified on KAPTIV-AETM. Top, crude hybridoma supernatant. Bottom, affinity purified IgA (peak 2). kDa pe cr ud e pe ak 1 pe ak 2 kDa SDS-PAGE analysis (silver staining) of mouse monoclonal IgA affinity purified on KAPTIV-AETM . Non reducing SDS-PAGE analysis (silver staining) of monoclonal IgE from ascites affinity purified on KAPTIV-AETM. A crude sample of ascitic fluid (1 ml) has been loaded on a KAPTIVAE™ column (2 ml) equilibrated at a flow rate of 1 ml/min with 50 mM sodium phosphate pH 7.0, at room temperature. Adsorbed IgE (recovery > 80 %) was eluted by a buffer change to 0.1 M acetic acid. 1 Purification of monoclonal IgE from ascites 0 5 10 15 Elution time [min] cr ud e simple use high IgA and IgE purity short purification time (20 minutes) adsorption in PBS or TRIS buffers high loading capacity (10 mg/ml matrix) elution of adsorbed antibodies in HAc 0.1 M, glycine 0.2 M pH 2.0, or sodium bicarbonate 0.1 M pH 9.0 broad selectivity for IgA and IgE from different species synthetic ligand, no biological contaminants full recovery of immunoreactivity stable to sanitizing agents stable to repeated use 66 Immunoreactivity recovery (%) ABS 280 nm - 0.01 A.U.F.S. Affinity purification of mouse monoclonal IgA from crude cell supernatant on KAPTIV-AETM. A 5 ml sample of crude supernatant has been loaded on a 2 ml KAPTIV-AE™ column (100 x10 mm I.D.) equilibrated at a flow rate of 1.0 ml/min with 100 mM sodium phosphate pH 7.0, at room temperature. After elution of the unbound material, the eluent has been changed to 0.1 M acetic acid. IgA recovery was >90% as determined by ELISA. peak 2 100 80 60 40 peak 1 20 0 KAPTIV-AETM Ready to use matrix for IgA and IgE purification Purification of IgA from serum samples, such as cell culture supernatants, ascites, or sera. Sample loading is achieved at room temperature with physiological buffers, and dissociation of bound immunoglobulins is achieved by a buffer change to 0.1 M acetic acid, 0.2 M glycine pH 2.2, or 0.1 M sodium bicarbonate pH 9.0, with full retention of immunological properties. Matrix capacity is at least 10 mg/IgA or IgE per ml of resin. KAPTIV-AE™ can be used in the batch mode or easily packed in columns for analytical as well as preparative applications, for a cost effective isolation of IgA or IgE with high purity degree. KAPTIV-AE™ is particularly useful for the down stream processing of monoclonal antibodies for therapy, given the high capacity and ligand stability. Recovery of IgA immunoreactivity from serum after affinity purification on KAPTIV-AE TM monitored by ELISA. A crude sample of mouse serum (0.2 ml) has been loaded on a KAPTIVAE™ column (2 ml) equilibrated at a flow rate of 1.0 ml/min with phosphate saline buffer (PBS) at room temperature. Adsorbed IgA was eluted by a buffer change to 0.1 M acetic acid and immediately neutralized. The majority of IgA immunoreactivity is found in the column adsorbed fraction (peak 2), while negligible activity is found in the column flow-through. Purification of monoclonal IgA from cell culture supernatants Immunoreactivity recovery (%) KAPTIV-AE™ is a new matrix optimized for the affinity purification of monoclonal or polyclonal IgA and IgE from crude peak 2 100 80 60 40 20 peak 1 0 peak 1 peak 2 190 First lane crude hybridoma supernatant. Second lane Unbound material from the KAPTIV-AE™ affinity column (peak 1) Third lane KAPTIV-AE™ affinity purified IgA (peak 2) ABS 280 nm - 0.002 A.U.F.S. Recovery of IgE immunoreactivity after affinity purification on KAPTIV-AETM. IgA crude IgA peak 2 0 Background: Structure of TG19318*, affinity ligand for IgA and IgE purification [*patents pending] 50 100 150 Elution time [min] 200 2 ak pe ak 190 First lane Crude ascitic fluid Second lane Column unretained material, only traces of IgE are visible (peak 1) Third lane KAPTIV-AE™ affinity purified IgE (peak 2) 66 serum albumin Gel permeation analysis of mouse monoclonal IgA affinity purified on KAPTIV-AETM. Top, crude hybridoma supernatant. Bottom, affinity purified IgA (peak 2). kDa pe cr ud e pe ak 1 pe ak 2 kDa SDS-PAGE analysis (silver staining) of mouse monoclonal IgA affinity purified on KAPTIV-AETM . Non reducing SDS-PAGE analysis (silver staining) of monoclonal IgE from ascites affinity purified on KAPTIV-AETM. A crude sample of ascitic fluid (1 ml) has been loaded on a KAPTIVAE™ column (2 ml) equilibrated at a flow rate of 1 ml/min with 50 mM sodium phosphate pH 7.0, at room temperature. Adsorbed IgE (recovery > 80 %) was eluted by a buffer change to 0.1 M acetic acid. 1 Purification of monoclonal IgE from ascites 0 5 10 15 Elution time [min] cr ud e simple use high IgA and IgE purity short purification time (20 minutes) adsorption in PBS or TRIS buffers high loading capacity (10 mg/ml matrix) elution of adsorbed antibodies in HAc 0.1 M, glycine 0.2 M pH 2.0, or sodium bicarbonate 0.1 M pH 9.0 broad selectivity for IgA and IgE from different species synthetic ligand, no biological contaminants full recovery of immunoreactivity stable to sanitizing agents stable to repeated use 66 Immunoreactivity recovery (%) ABS 280 nm - 0.01 A.U.F.S. Affinity purification of mouse monoclonal IgA from crude cell supernatant on KAPTIV-AETM. A 5 ml sample of crude supernatant has been loaded on a 2 ml KAPTIV-AE™ column (100 x10 mm I.D.) equilibrated at a flow rate of 1.0 ml/min with 100 mM sodium phosphate pH 7.0, at room temperature. After elution of the unbound material, the eluent has been changed to 0.1 M acetic acid. IgA recovery was >90% as determined by ELISA. peak 2 100 80 60 40 peak 1 20 0 KAPTIV A product line designed for the affinity purification of monoclonal or polyclonal antibodies from different sources. Ligands are synthetic peptidomimetic molecules, stable to repeated use and ideal for analytical and preparative uses. Selective matrices are available for IgG, IgM, IgA, IgE and IgY purification KAPTIV-AETM is available directly from TECNOGEN s.c.p.a. in different formats product code description TG9001 TG9002 TG9003 TG9004 KAPTIV-AETM KAPTIV-AETM KAPTIV-AETM KAPTIV-AETM KAPTIV-AE TM Af f inity matrix f o r I g A and IgE p ur i f i c a t i o n (5 ml) (10 ml) (25 ml) (100 ml) KAPTIV-AETM is available also in bulk amounts for preparative applications. Please contact our sales department for quotations and additional products of the KAPTIV line. KAPTIV-MTM Affinity matrix for IgM purificaiton KAPTIV-M product code description TG8001 TG8002 TG8003 TG8004 KAPTIV-MTM KAPTIV-MTM KAPTIV-MTM KAPTIV-MTM ™ (5 ml) (10 ml) (25 ml) (100 ml) to order Orders may be placed by phone, fax or E-mail directly to: TECNOGEN s.c.p.a. Tecnogen S.p.A. Parco Scientifico, Località La Sede legale: Località LaFagianeria Fagianeria - Piana di 81015 Piana di Monte Verna (CE) - ITALY Monte Verna (CE) - Italia Phone: +39-0823-612-208/228, Fax: +39-0823-612-222/230 Reception: +39 0823 612 301 E-mail: [email protected] Home page: http://www.tecnogen.it Capture of IgA and IgE made easy TECNOGEN s.c.p.a.
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