microDAWN

microDAWN

The Only Multi-Angle Light Scattering Detector for UHPLC.

UHPLC-SEC System

The microDAWN™ can be coupled to any UHPLC-SEC system to determine absolute molecular weights and sizes of polymers, peptides, or proteins directly, without resorting to column calibration or reference standards. The microDAWN produces minimal band broadening in order to maintain the narrow peaks typical of UHPLC, while meeting or exceeding the sensitivity of the miniDAWN.


Polymer Characterization

Read our paper published in Nature Scientific Reports: “Measuring proteins with greater speed and resolution while reducing sample size”


Use of SEC-MALS

Download our white on the microDAWN:“Absolute characterization of polymers with light scattering and UHP-SEC”

The microDAWN MALS detector performs absolute characterization of macromolecules eluting from UHPLC, determining molar mass and size independently of retention time and molecular reference standards. This instrument offers superb sensitivity over a wide range of molar mass, size and concentrations. In most respects the microDAWN is similar to the well-established miniDAWN , a workhorse MALS instrument used in hundreds of labs around the world for characterizing proteins and other molecules smaller than 50 nm in radius.

Distinctly from the miniDAWN, the microDAWN is uniquely suitable for use in conjunction with UHPLC Size Exclusion Chromatography (SEC-MALS) thanks to its small interdetector dispersion. In some cases it may be used with ion-exchange or other types of chromatography. Regardless of the specifics, the microDAWN produces minimal band broadening in order to maintain the narrow peaks typical of UHPLC.

microDAWN UHPLC Experimental Setup

  • Mw from 200 Da to 10 MDa
  • Sensitivity: 1.0 µg/mL BSA (66.4 kDa) in PBS; 50 ng/mL 100 kDa polystyrene in THF.
  • Rg from 10 to 50 nm
  • Ultra-stable laser

The microDAWN and its companion microOptilab Refractive Index Detector interface to most standard UHPLC systems. As these detectors are based on the advanced technology of Wyatt's miniDAWN and Optilab, respectively, users may be confident in the underlying capabilities cited in thousands of peer-reviewed publications.

Ultrasonic Cleaning Included

Each microDAWN includes a COMET ultrasonic transducer that cleans MALS flow cells automatically by agitating the fluid and loosening particles from the glass surfaces.

microDAWN UHPLC Experimental Setup

The microDAWN will work downstream of all types of UHPLC systems for analysis of molar mass and size distributions (see SEC-MALS), and derivatives such as conformational analysis. For those systems equipped with a UV, RI, fluorescence or other type of concentration detector, the microDAWN accepts up to 4 analog inputs so a 3rd-party concentration signal may be used in MALS calculations of molar mass. The microDAWN is set up downstream of a UV detector and upstream of a RI detector.

In most cases it is beneficial to connect an microOptilab as well as a UV detector. This permits universal concentration detection regardless of fluorophores or chromaphores. The combination of UV and RI signals is utilized in ASTRA's Conjugation Analysis algorithm to characterize binary complexes such as glycoproteins, PEGylated proteins, membrane proteins enveloped in surfactant, or block co-polymers.

WyattQELS DLS Module - To determine size and conformation, the WyattQELS measures hydrodynamic radius of eluting macromolecules in the same flowcell as the MALS detectors. This fully integrated module guarantees dispersionless correlation between molar mass, rms radius and hydrodynamic radius.

Alternatively, connect an external DLS instrument via fiber optic cable - both the DynaPro NanoStar and Mobius can do double duty as batch and online DLS instruments. Since they collect light directly from the MALS flow cell, the mass/size correlation is still dispersionless.

ASTRA - Our comprehensive software solution for MALS and DLS analysis in chromatography, AF4 or batch mode. ASTRA is available in a 21 CFR(11) compliant version and offers additional options such as particle analysis and a research database.

ASTRA's HPLC Control service allows for full digital synchronization between your UHPLC system.

ASTRA's Method Wizard lets you set up a default method optimized for your sample type in three easy steps:

  1. Select experiment type
  2. Input parameters
  3. Hit Run

Once the method is ready, you are just a click away from MALS analysis with One-Click Mw™. ASTRA will:

  1. Synchronize data collection with your UHPLC
  2. Autoset parameters to determine MW and
  3. Rg
  4. Generate custom reports and graphs
  5. Prepare for the next run
HPLC Manager
Measurements  
   Molar Mass Range 200 Da to 10 MDa1
   Molecular or Particle Size Range
   (Radius, Rg)
10 nm – 50 nm 1, up to 150 nm with shape-specific models
   Molecular Size Range (Rh, with    WyattQELS DLS module or
   DynaPro NanoStar)
0.5 to 30 nm, flow rate 0.3 mL/min1
0.5 to 20 nm, flow rate of 0.5 mL/min1
   Sensitivity 3.5 ng of 100 kDa polystyrene in THF2
70 ng of BSA in PBS2
Sample Temperature Control Ambient
Optics  
Detectors  
   MALS Detectors High-gain, high dynamic range, static light scattering detectors at 3 angles
   Auxiliary Detectors Laser monitor for stabilization feedback;
Forward transmission monitor to correct signals for absorbing samples and to assess data quality.
   A/D Resolution 24 bit (detector dynamic range > 16,000,000)
Sample Temperature Control ambient only
Laser Properties
   Laser Power Control Programmable 10% - 100%
   Wavelength λ = 660 nm (nominal). Each instrument is individually characterized.
Band Broadening Upstream detector to microDAWN: < 1.5 μL;
microDAWN to downstream detector: <1.6 μL FWHM @ 0.3 mL/min
Wetted Materials 316 stainless steel, fused silica, MP35N, PEEK, PTFE
Integrated Cell Cleaning Embedded COMET flow cell cleaning system is included with every microDAWN
Instrument Options Embedded WyattQELS for simultaneous DLS measurements
   
Electronics  
   Analog Inputs 4 differential analog inputs with 24 bit resolution
   Analog Outputs 3 single ended analog outputs -10 V to +10 V
   Other Inputs/Outputs Alarm in, Autoinject contact closure, Recycle in, Autoinject retransmit, Alarm retransmit, Recycle valve power
   Front Panel Display 162.5 mm, 16-bit, high resolution touch screen + physical keypad
   Computer Interface Ethernet
   Processed Data Transmission Rate 73 Hz to 0.2 Hz
Dimensions 60 cm (L) x 36 cm (W) x 17 cm (H)

1Typical. Actual range depends on dn/dc, sample concentration, sample conformation and chromatography conditions.

2Under UHPLC-SEC conditions with 4.6x150 mm SEC columns.


Host PC requirements may be found in Computer Requirements.

Specifications subject to change without notice.

"Wonderful instrument, supports daily activities."
"Advances in UHPLC technology have greatly reduced run times, sample loading requirements, and solvent consumption. The µDAWN seamlessly integrates downstream from our Agilent 1290 UHPLC for accurate measurements of Mw, Rz and Rh values, giving us a deeper look into our sample beyond just the UV chromatogram. It is just as easy to use as our other Wyatt MALS detectors and is used almost daily." William Wittbold, Ajinomoto Althea

Other MALS Instruments

 dawn, molar mass determination, molar mass measurement, molar mass analysis, molecular weight measurement, molar mass characterization

DAWN® - The most sensitive MALS detector available, anywhere. Incorporates detectors at 18 angles to determine molar masses from 200 Da to 1 GDa and radii from 10 – 500 nm.

  • Standard option: ambient temperature
  • Heated/cooled option: -15°C to +150°C
  • High-temperature option: ambient to +210°C

The DAWN offers special options to handle fluorescent samples: fluorescence-blocking filters and an infrared, 785 nm laser.

miniDAWN® - Second only to the DAWN in sensitivity. Incorporates detectors at 3 angles to determine molar masses from 200 Da to 10 MDa and radii from 10 – 50 nm. Ambient only.

ultraDAWN™ - Measures multi-angle light scattering and reports the results – weight-average molecular weight Mw and rms radius Rg – in real time.