Nexera Bio Biocompatible Liquid Chromatograph
Nexera Bio is a biocompatible UHPLC system offering the same superior reliability, robustness, and expandability as our flagship Nexera UHPLC systems. But, Nexera Bio is distinctive in that it is well suited to the analysis of protein-based pharmaceuticals, antibody drugs, and other substances related to biotechnologies, genetic engineering, cell fusion, and cell culturing.
Nexera Bio is compatible with mobile phases containing high concentrations of salts or acids typically used for analyzing antibody drugs and membrane proteins. Its low surface activity inhibits peak tailing experienced on traditional UHPLC systems. Such tailing can be especially problematic when analyzing phosphorylated peptides, which have a high affinity for metals.
Nexera Bio Features
1. Low Carryover
Shimadzu autosamplers have a reputation for the lowest carryover in the industry. Nexera Bio features specially selected materials and a rinsing mechanism that further inhibit carryover from biological samples. Peak detection at low levels and chromatographic reproducibility are dramatically improved.
2. Minimized Sample Loss due to Adsorption
Some biological molecules adsorb readily to stainless steel tubing. Successful assays for such samples require a system with low surface activity to minimize this problem. The low surface activity of Nexera Bio results in minimized sample loss and greatly improved sensitivity.
3. Improved Corrosion Resistance
The analyses of biological samples frequently require mobile phases with high salt concentrations, halogenated compounds or ion pairing agents. These conditions can be problematic for stainless steel UHPLC systems. With Nexera Bio, crucial metal-free components define the wetted surfaces while maintaining a high-efficiency flow path (66 MPa). The system is unaffected by high salt content (KH2PO4 in H3PO4), and the corrosion resistant design provides trouble-free operation.
2. Nexera Bio Advantages
2-1. High-Sensitivity Analysis
The figure to the right shows a chromatogram obtained from flow injection analysis of an adenosine triphosphate (ATP) standard solution (1 mg/L) using Nexera Bio and a general-purpose UHPLC system.
Because ATP exhibits strong affinity for metals, analysis using a general-purpose UHPLC is difficult because of analyte binding in the system. By contrast, Nexera Bio inhibits such adsorption, making good peak shapes and high sensitivity a reality.
|1st Inj.||2nd Inj.||3rd Inj.|
2-2. Excellent Reproducibility
The figure to the right shows a comparison of peak area values from 10 successive ATP analyses (standard solution at 1 mg/L) using Nexera Bio and general-purpose UHPLC systems.
Because of its low surface activity, Nexera Bio achieves much higher sensitivity and better reproducibility than the general-purpose UHPLC system.
|Mean Value（n=10）||Coefficient of Variation（%）|
3. Consumables Ideal for the Analysis of Biopharmaceuticals
Shim-pack Bio Diol and IEX columns offer performance that can help increase the accuracy of analyzing and evaluating biopharmaceuticals and drugs based on mid-sized molecules.
3-1. Example Using Shim-pack Bio Diol Columns for Evaluating Aggregates in Antibody Drugs
In this example, Shim-pack Bio Diol water-based size exclusion chromatography columns were used to evaluate aggregates in an antibody drug.
Available in four different pore sizes, Shim-pack Bio Diol columns are suitable for size exclusion chromatography and intended for bioanalysis. They are ideal for analyzing aggregates or separating fragments in antibody drugs, separating nucleic acids or sugar chains, and for measuring molecular weight.
The figure below shows an example using a Shim-pack Bio Diol column to analyze an antibody drug.
Reducing the particle size from 5 µm to 2 µm improved aggregate separation and also resulted in higher sensitivity and quantitative accuracy. Furthermore, high-speed analysis was achieved by using a shorter (150 mm long) column.
Increasing the Speed of Antibody Drug Aggregate Analysis by Using a Column with 2 µm Particle Size
3-2. Example Using a Shim-pack Bio IEX Column for Synthetic Oligonucleotides
The Shim-pack Bio IEX is an ion-exchange column suitable for separation of proteins and nucleic acids. Shim-pack Bio IEX columns are available with either non-porous or porous particles and are intended for laboratory-scale purification of small quantities.
For the figure below, a non-porous Shim-pack Bio IEX column was used to analyze synthetic oligonucleotides. The Shim-pack Bio IEX is able to distinguish between molecules that differ by only one salt base.
Separation of Synthetic Oligonucleotide (Single-Chain DNA) Using a Shim-pack Q-NP Column
3-3. Example of Peptide Analysis Using a Mastro™ C18 Metal-Free Column
The chromatogram shown below was obtained by successive analysis of a standard solution of angiotensin I (100 mg/L; n=5) using a Mastro C18 high-pressure metal-free column in a Nexera Bio system.
The 0.12 % relative standard deviation (n = 5) in peak area values indicates excellent reproducibility.
|Mean Value （n=5）||Coefficient of Variation（%）||Symmetry Factor|
Nexera Bio system combined with a Mastro C18 column achieved both low adsorption and high sensitivity, which should help improve the reliability of biopharmaceutical analysis.
4. Column Product Line
4-1. Shim-pack Bio Diol
|Shim-pack Bio Diol||Diol-60||Diol-120||Diol-200||Diol-300|
|Functional Group||Dihydroxypropyl (Diol)|
|Particle Size||3 µｍ，5 µm||2 µm，3 µm，5 µm|
|Pore Size||6 nm||12 nm||20 nm||30 nm|
|pH Range Used||5.0 - 7.5|
|Target Molecular Weight Range||10,000 or less||1,000 to 100,000||5,000 to 300,000||20,000 to 1,000,000|
|Particle Size||2 µm||3 µm|
|4.6 x 150 mm||227-31009-01||227-31010-01|
|4.6 x 300 mm||227-31009-02||227-31010-02||227-31007-01||227-31008-01||227-31009-03||227-31010-03|
|Pressure Capacity||45 MPa||20 MPa|
|Particle Size||5 µm|
|4.6 x 300 mm||227-31007-02||227-31008-02||227-31009-04||227-31010-04|
|8.0 x 300 mm||227-31007-03||227-31008-03||227-31009-05||227-31010-05|
|8.0 x 30 mm (Guard Column)||227-31007-04||227-31008-04||227-31009-06||227-31010-06|
|Pressure Capacity||20 MPa|
|20 x 300 mm||227-31097-01||227-31098-01||227-31099-01||227-31100-01|
|20 x 500 mm||227-31097-02||227-31098-02||227-31099-02||227-31100-02|
|20 x 50 mm (Guard Column)||227-31116-01||227-31117-01||227-31118-01||227-31119-01|
|Pressure Capacity||10 MPa|
|Shim-pack Bio IEX||Q-NP||SP-NP||Q||SP|
|Substrate||Hydrophilic non-porous polymer||Hydrophilic porous polymer|
|Functional Group||- CH2N+(CH3)3||- (CH2)3SO3-||- CH2N+(CH3)3||- (CH2)3SO3-|
|Particle Size||3 µｍ，5 µm||5 µm|
|pH Range Used||5.0 - 7.5|
|Shim-pack Bio IEX||Q-NP||SP-NP||Q||SP|
|3 µm||5 µm||3 µm||5 µm||5 µm|
|4.6 x 30 mm||227-31002-01||227-31003-01||227-31005-01||227-31006-01||227-31001-01||227-31004-01|
|4.6 x 50 mm||227-31002-02||227-31003-02||227-31005-02||227-31006-02||227-31001-02||227-31004-02|
|4.6 x 100 mm||227-31002-03||227-31003-03||227-31005-03||227-31006-03||227-31001-03||227-31004-03|
4-3. Mastro C18 High-Pressure Metal-Free Column
Peak tailing due to system adsorption is common when analyzing compounds containing a phosphate group, such as phosphorylated peptides and nucleotides. Mastro™ series columns are high-pressure, metal-free columns that feature a polymer-lined stainless steel column body and highly pressure-resistant polymeric frit. Two models are available in the Mastro series, the Mastro™ C18 and the Mastro™ PFP2 (pentafluorophenyl).
The Mastro™ PFP2 (pentafluorophenyl) is a reversed-phase column suitable for separating compounds that are difficult to retain on many C18 columns. Examples are positional isomers and some halogenated compounds.
The figure to the right compares results obtained from a standard solution of ATP (10 mg/L) in a Nexera Bio system using a general-purpose ODS column (stainless steel column) and a Mastro C18 high-pressure metal-free column.
The ATP could not be detected using the general-purpose ODS column, due to adsorption to the metal surfaces inside the column, but the ATP was detected without any sample loss using the polymer-lined Mastro C18. These results show how the Nexera Bio system in combination with a Mastro C18 column can be used to reliably analyze nucleotides and other compounds that include a phosphate group.
|Mean Value（n=3）||Coefficient of Variation(%)|
|SUS Body C18||N.D.||-|
>> For inquiries regarding Mastro columns, click here (Shimadzu GLC).
5. Nexlock PLS High-Pressure PEEK Tubing and Finger-Tightened Fittings
Nexera Bio systems use highly pressure-resistant PEEK tubing (Nexlock PLS*), reinforced with stainless steel, to connect the autosampler and column. The fittings seal the tubing connection in such a way that minimizes dead volume helps to prevent peak tailing. Because they are finger-tight, no tools are needed.
* 66 MPa pressure capacity indicated in specifications, with replacement recommended after 100 uses
6. TORAST H-Bio Vial
Adsorption Test of Myoglobin Trypsin Digest Products
Reportedly, the main mechanisms for adsorption of peptides to vials are hydrophobic adsorption and ionic adsorption. With glass vials, ionic adsorption to silanol groups at the glass surface dominates. With polymeric vials, hydrophobic adsorption dominates. Shimadzu GLC has developed a polymeric vial with a special coating that minimizes the adsorption of peptides. Results have indicated that the vial is inert to basic, acidic, and neutral compounds. It is especially recommended for LC and LC/MS(/MS) users that frequently analyze low-concentration samples.