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How FPLC Systems Work & How Leasing One Can Benefit Your Lab

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Fast protein liquid chromatography (FPLC) diagram

Fast protein liquid chromatography (FPLC) is a medium- or low-pressure liquid chromatography method for purifying, and occasionally analyzing, different mixtures of large biomolecules.

Chromatography diagram

It is also used to collect separated compounds of interest and identify protein profiles within single Biomolecule mixtures can include proteins, peptides, oligonucleotides, DNA, and RNA. Because biomolecules are highly sensitive and can’t withstand the type of high pressure, temperature, or solvents used in high-performance liquid chromatography (HPLC), FPLC is employed. FPLC is also used to collect separated compounds of interest and identify protein profiles within single proteins.

You may hear FPLC referred to as medium-pressure chromatography, biochromatography, or biopurification, but the title doesn’t change the process. It’s an important method for many researchers in the life sciences and biotech.

Separating a sample within a mixture depends on each molecule’s specific interaction with the stationary and mobile phases used during chromatography. FPLC utilizes a wide range of resins, composed of many small coated beads, for its stationary phase, and a an aqueous solution or buffer for its mobile phase.

The targeted protein adheres to the resin as it flows through the separation column and the buffer carries any other molecules that are present out of the system. It’s typical to see a mixture of buffers used to accomplish this elution.

For example, having flushed the system, a secondary buffer, called an eluent, will separate the protein of interest from the resin and pull it into a detection system for measurement, recording the concentration levels of the target protein in the sample.

FPLC is a type of preparative technique, unlike HPLC which provides quantitative and qualitative analysis of liquid samples. It is used in all sorts of settings, from academia to research facilities to industrial manufacturing.

Its versatility is due in part to its simplicity, rapidity, and reliability as a technique, producing consistent results that end-users can count on.

Furthermore, its ability to support multiple columns simultaneously is based on its use of low pressure and offers a wide flow range when combined with multiple pumps, making the entire process efficient and highly scalable.

Some of the most common FPLC systems on the market include the AKTA FPLC system, offered by Cytiva—formerly GE Healthcare and Amersham Biosciences—and the AZURA® FPLC system, offered by KNAUER. (And guess what? We lease them.)

FPLC System Components, Modes, & Price

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Fast protein liquid chromatography chromatographs operate using several key components and chromatographic modes.

Components

FPLC systems consist of the following features:

  • Program controller: This controls many aspects of the system, from separation parameters to data collection.
  • Pump: It is typical to see two pumps used to control the amount of each buffer introduced during liquid chromatography. This is often a type of piston pump. Occasionally, a single pump will be used where the buffers are drawn from their separate reservoirs into a proportioning valve attached to a mixing chamber.
  • Injection valve: This valve connects the sample loop and mixer to the column. The valve is able to load the sample into the injection loop, which injects the sample into the column.
  • Injection loop: This tubing is filled with the sample solution before it is injected into the column.
  • Column: Made of glass or plastic, this chromatographic column is longer than one typically used during HPLC, but has a large internal diameter, as the samples used in FPLC are larger. The column is packed with the resin and buffer solution, with a small opening on one end to retain the beads while letting the dissolved proteins and buffer pass through.
  • Flow cells: The eluent passes through one or more flow cells which measure the concentration of protein in the eluent using UV light, salt concentration using conductivity, and even measure the level of pH. These monitors allow for detection of proteins when individually present in the eluent.
  • Fraction Collector: This component is made up of small containers such as test tubes that collect the eluent in fixed volumes or specific fractions based on peak protein concentration.
  • Monitor: The monitor, or recorder, consists of a computer and interface with a display. It identifies and records when concentrations of biomolecules are present.

Ion Exchange

This method separates molecules based on their specific charge, which depends on the pH or buffer. If a protein is more charged, it will be more likely to bind with an oppositely charged resin. Depending on salt concentration, a protein will take shorter or longer to elute from the resin as the salt ions compete with the molecule for adherence to the resin.

Affinity

Using a substrate that is linked with the stationary phase, a sample is passed through and binds to the solid substrate according to its affinity for the substrate used. The compounds that haven’t bound are then washed out using an aqueous solution. The remaining compounds that are bound to the substrate are then eluted using a solvent with a certain pH or salt concentration.

Size Exclusion

This method is also referred to as gel filtration chromatography. A gel medium that consists of specifically sized beads that separate the sample mixture as it passes through. Molecules that are larger cannot pass through and are eluted from the column, while the smaller molecules diffuse into the gel medium and remain fixed in place. This results in separation based on size and molecular weight, as elution happens in order of linear weight.

We Offer Biochromatography Leases to Fit Every Need

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Founder-Friendly Leases

Our lease agreements are founder-friendly and flexible, helping you preserve working capital, strengthen the cash flow of your business, and keep business credit lines open for expansions, staffing, and other crucial operational expenses and business development opportunities.

2-5 Year Lease Lengths

Leases range from 2 to 5 years. Length will depend on several factors, including how long you want to use the equipment, equipment type, and your company’s financial position. These are standard factors leasing companies consider and help us tailor a lease agreement to fit your needs.

Your Choice of Manufacturer

We don’t carry an inventory. This means you’re not limited to a specific set of manufacturers. Instead, you can pick the equipment that aligns with your business goals and preferences. We’ll work with the manufacturer of your choice to get the equipment in your facility as quickly as possible.

Maintenance & Repair Coverage

Bundle preventive maintenance and repair coverage with your lease agreement. You can spread those payments over time. Easily maintain your equipment, minimize the chances something will break down, repair instrumentation quickly, and simplify your payment processes.

End-of-Lease Options

At the end of your lease, you have multiple options. You can either renew the lease at a significantly lower price, purchase the machine outright based on the fair market value of the original pricing, or call it a day and we’ll come the pick up the equipment for you free of charge.

No Loan-Like Terms

Our leases do not include loan-like terms, which can be restrictive or harmful in certain situations. We do not require debt covenants, IP pledges, collateral,  or equity participation. Our goal is to maximize your flexibility. When you lease with us, you’re collaborating with a true business partner.

In-House Underwriting Process

Our underwriting is done in-house. You can expect quicker turnaround, allowing you respond to your equipment needs as they arise. We require less documentation than traditional lenders and financiers and can get the equipment you need in operation more quickly.

Popular FPLC System Manufacturers

Bio-Rad
Cytiva
Knauer