Online coupling of analytical hydrophobic interaction chromatography with native mass spectrometry for the characterization of monoclonal antibodies and related products.
The original mass spectrometry (MS native) has seen a remarkable development and improvement of applications over the last decade to the study of proteins and protein complexes. Although conventionally carried out using a static nanospray emitter in offline mode, the original MS has been increasingly applied in hyphenated methods, in which various separation techniques are directly coupled to the genuine online MS detection. Those new developments have greatly expanded the usefulness of the original MS characterization of protein biopharmaceuticals. analytical methods hydrophobic interaction chromatography (HIC), although it is often used for the characterization of monoclonal antibodies (mAbs) and antibody-drug conjugate (ADC), rarely explored for online coupling with the original MS.
This is largely due to the high salt concentration used in HIC analysis that is not compatible with MS detection directly. In this study, we address this challenge through makeup and flow splitting innovative design and successfully achieve coupling online analytical HIC separation with the original MS detection. Development and experimental setup HIC-MS method is described in detail to explain how this design can cope with high salt concentrations used in HIC separation and ultimately to achieve both a good chromatographic resolution and data quality MS.
Furthermore, the utility of HIC-MS method is shown in three different applications, where a mixture of mAb, mAb variant molecule resulting from the PTM, and imitate all the Cys-based ADC is characterized in detail. Unlike previously reported HIC-MS method, this new method was developed using analytical HIC column scale with conventional ligand so that the separation profile is achieved very comparable to those obtained with standard methods of HIC-UV. Consequently, HIC-MS method is not only provide an alternative approach to in-depth characterization of mAbs and related products during their development, but also can be applied immediately to help peak assignment and identity explanation for HIC-UV method is used in quality control.
Online coupling of analytical hydrophobic interaction chromatography with native mass spectrometry for the characterization of monoclonal antibodies and related products.
Plasma Donors in the Southwestern United States Positive Contribution to Profile Multi Immune Globulin Therapeutic Antibody Products.
Human plasma-derived immune globulin (IG) used in augmentation therapy to provide the level of protection of the antibody for patients with primary immune deficiency disease (PIDD) and for prophylaxis against infectious diseases.
To keep the extent of antibodies needed for clinical protection, it is important to understand the pattern of areas with a prevalence of antibodies in the plasma source from which the IG product is produced. In this study, plasma source of a donation center at various locations on the US West quarter surveyed for antibody titers against hepatitis A virus (HAV), measles virus (MeV), and cytomegalovirus (CMV). A variety of anti-HAV Ig titer plasma was observed between the centers, with some centers exhibiting titer another 3-5 times.
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human Slitrk4 . This antibody is tested and proven to work in the following applications:
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human Slitrk4 . This antibody is tested and proven to work in the following applications:
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human SLITRK4 (C-term). This antibody is tested and proven to work in the following applications:
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human SLITRK4 (N-Terminus). This antibody is tested and proven to work in the following applications:
Description: Slitrk4 Antibody: SLIT and NTRK-like family 4 (Slitrk4) is a member a protein family consisting of six homologous transmembrane proteins (Slitrk1-6) that share two conserved leucine-rich repeat domains in the extracellular domain and have significant homology to Slit, a secreted axonal growth-controlling protein. These proteins are also homologous to trk neurotrophin receptors in their intracellular domains. Expression of Slitrk proteins is highly restricted to neural and brain tumor tissues, but varies within the protein family. Like every other Slitrk protein except Slitrk1, overexpression of Slitrk4 inhibited neurite outgrowth in cultured neurons, suggesting that these proteins are involved in the control of neurite outgrowth.
Description: Slitrk4 Antibody: SLIT and NTRK-like family 4 (Slitrk4) is a member a protein family consisting of six homologous transmembrane proteins (Slitrk1-6) that share two conserved leucine-rich repeat domains in the extracellular domain and have significant homology to Slit, a secreted axonal growth-controlling protein. These proteins are also homologous to trk neurotrophin receptors in their intracellular domains. Expression of Slitrk proteins is highly restricted to neural and brain tumor tissues, but varies within the protein family. Like every other Slitrk protein except Slitrk1, overexpression of Slitrk4 inhibited neurite outgrowth in cultured neurons, suggesting that these proteins are involved in the control of neurite outgrowth.
Description: Slitrk4 Antibody: SLIT and NTRK-like family 4 (Slitrk4) is a member a protein family consisting of six homologous transmembrane proteins (Slitrk1-6) that share two conserved leucine-rich repeat domains in the extracellular domain and have significant homology to Slit, a secreted axonal growth-controlling protein. These proteins are also homologous to trk neurotrophin receptors in their intracellular domains. Expression of Slitrk proteins is highly restricted to neural and brain tumor tissues, but varies within the protein family. Like every other Slitrk protein except Slitrk1, overexpression of Slitrk4 inhibited neurite outgrowth in cultured neurons, suggesting that these proteins are involved in the control of neurite outgrowth.
Description: Slitrk4 Antibody: SLIT and NTRK-like family 4 (Slitrk4) is a member a protein family consisting of six homologous transmembrane proteins (Slitrk1-6) that share two conserved leucine-rich repeat domains in the extracellular domain and have significant homology to Slit, a secreted axonal growth-controlling protein. These proteins are also homologous to trk neurotrophin receptors in their intracellular domains. Expression of Slitrk proteins is highly restricted to neural and brain tumor tissues, but varies within the protein family. Like every other Slitrk protein except Slitrk1, overexpression of Slitrk4 inhibited neurite outgrowth in cultured neurons, suggesting that these proteins are involved in the control of neurite outgrowth.
Description: A polyclonal antibody against SLITRK4. Recognizes SLITRK4 from Human. This antibody is Unconjugated. Tested in the following application: ELISA, IHC; Recommended dilution: IHC:1:20-1:200
Description: A polyclonal antibody against SLITRK4. Recognizes SLITRK4 from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, WB
Description: A polyclonal antibody against SLITRK4. Recognizes SLITRK4 from Human, Mouse. This antibody is Unconjugated. Tested in the following application: ELISA, IHC;ELISA:1:1000-1:2000, IHC:1:10-1:50
Description: A polyclonal antibody against SLITRK4. Recognizes SLITRK4 from Human, Mouse. This antibody is Unconjugated. Tested in the following application: ELISA, WB, IHC;ELISA:1:1000-1:2000, WB:1:200-1:1000, IHC:1:10-1:50
Description: Members of the SLITRK family, such as SLITRK5, are integral membrane proteins with 2 N-terminal leucine-rich repeat (LRR) domains similar to those of SLIT proteins (see SLIT1; MIM 603742). Most SLITRKs, including SLITRK5, also have C-terminal regions that share homology with neurotrophin receptors (see NTRK1; MIM 191315). SLITRKs are expressed predominantly in neural tissues and have neurite-modulating activity (Aruga et al., 2003 [PubMed 14557068]).
Description: Members of the SLITRK family, such as SLITRK5, are integral membrane proteins with 2 N-terminal leucine-rich repeat (LRR) domains similar to those of SLIT proteins (see SLIT1; MIM 603742). Most SLITRKs, including SLITRK5, also have C-terminal regions that share homology with neurotrophin receptors (see NTRK1; MIM 191315). SLITRKs are expressed predominantly in neural tissues and have neurite-modulating activity (Aruga et al., 2003 [PubMed 14557068]).
Description: Members of the SLITRK family, such as SLITRK5, are integral membrane proteins with 2 N-terminal leucine-rich repeat (LRR) domains similar to those of SLIT proteins (see SLIT1; MIM 603742). Most SLITRKs, including SLITRK5, also have C-terminal regions that share homology with neurotrophin receptors (see NTRK1; MIM 191315). SLITRKs are expressed predominantly in neural tissues and have neurite-modulating activity (Aruga et al., 2003 [PubMed 14557068]).
Description: Members of the SLITRK family, such as SLITRK5, are integral membrane proteins with 2 N-terminal leucine-rich repeat (LRR) domains similar to those of SLIT proteins (see SLIT1; MIM 603742). Most SLITRKs, including SLITRK5, also have C-terminal regions that share homology with neurotrophin receptors (see NTRK1; MIM 191315). SLITRKs are expressed predominantly in neural tissues and have neurite-modulating activity (Aruga et al., 2003 [PubMed 14557068]).
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human Slitrk1 . This antibody is tested and proven to work in the following applications:
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human Slitrk1 . This antibody is tested and proven to work in the following applications:
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human Slitrk2 . This antibody is tested and proven to work in the following applications:
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human Slitrk2 . This antibody is tested and proven to work in the following applications:
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human Slitrk3 . This antibody is tested and proven to work in the following applications:
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human Slitrk3 . This antibody is tested and proven to work in the following applications:
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human Slitrk5 . This antibody is tested and proven to work in the following applications:
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No small differences were observed for the level of anti-MeV and anti-CMV, respectively. Importantly, the increase in anti-HAV Ig titers were widely observed in all units of plasma obtained from centers showing high titer, indicative of a potential regional phenomenon among donors as opposed to some donors with high titers single. Plasma from the center of high-titer is given significantly greater neutralization against HAV in vitro. The results of this study provide a glimpse of antibody diversity inherent in human plasma that is used to produce IG
Evelyn
