Nexera Bio

 

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. 



 

For inquiries regarding TORAST H-Bio vials, click here (Shimadzu GLC).
Shimadzu GLC Ltd. 5F CS-Tower Bldg.,5-20-8, Asakusa-Bashi,Taito-ku,Tokyo 111-0053, Japan
E-mail : gsupport@glc.shimadzu.co.jp

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


Instrument Sensitivity Comparison for ATP 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.
Nexera Bio 6,242 6,357 6,221
General-PurposeUHPLC 825 1,121 1,389

 

 

2-2. Excellent Reproducibility


Instrument Reproducibility Comparison for ATP Analysis

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(%)
Nexera Bio
6,390
1.8
General-PurposeUHPLC 1,177 13.0
 

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.

The Analysis of Angiotensin I (n = 5)
Using a Nexera Bio System and Mastro Column



 
Mean Value (n=5) Coefficient of Variation(%) Symmetry Factor
416,572 0.12 % 1.2

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
Substrate Silica gel
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
 
 
 

Ordering Information

Particle Size 2 µm 3 µm
Packing Material Diol-200 Diol-300 Diol-60 Diol-120 Diol-200 Diol-300
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
Packing Material Diol-60 Diol-120 Diol-200 Diol-300
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
 
 


 

4-2. Shim-pack Bio IEX

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
 
 
 

Ordering Information

Shim-pack Bio IEX Q-NP SP-NP Q SP
Porosity Non-porous Porous
  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

Cut-Away View of a Mastro 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.

 

Effectiveness of Mastro C18 Column in Reducing Adsorption in ATP Analysis

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(%)
Mastro™ C18 38,481 4.5
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.

For inquiries regarding TORAST H-Bio vials, click here (Shimadzu GLC).
Shimadzu GLC Ltd. 5F CS-Tower Bldg.,5-20-8, Asakusa-Bashi,Taito-ku,Tokyo 111-0053, Japan
E-mail : gsupport@glc.shimadzu.co.jp

For Research Use Only. Not for use in diagnostic procedures.

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