Bioanalytical Technique Practical

[Protein Standard] (µg/ml)

Dilution factor

Volume Protein Standard (µl)

What’s the Price for a Paper?

Starts at $10/page for undergrad, up to $21 for pro-level. Deadlines (3 hours to 14 days) and add-ons like VIP support adjust the cost. Discounts kick in at $500+—save more with big orders!

Volume dH2O (µl)

Final volume (µl)

0

0

Is My Privacy Protected?

100%! We encrypt everything—your details stay secret. Papers are custom, original, and yours alone, so no one will ever know you used us.

0

150

200

25

Is AI Involved in Writing?

Nope—all human, all the time. Our writers are pros with real degrees, crafting unique papers with expertise AI can’t replicate, checked for originality.

1 in 80

2.5

197.5

200

Why Are You the Best for Research?

Our writers are degree-holding pros who tackle any topic with skill. We ensure quality with top tools and offer revisions—perfect papers, even under pressure.

125

1 in 16

12.5

187.5

Who Writes My Assignments?

Experts with degrees—many rocking Master’s or higher—who’ve crushed our rigorous tests in their fields and academic writing. They’re student-savvy pros, ready to nail your essay with precision, blending teamwork with you to match your vision perfectly. Whether it’s a tricky topic or a tight deadline, they’ve got the skills to make it shine.

200

250

1 in 8

25

Will My Paper Be Unique?

Guaranteed—100%! We write every piece from scratch—no AI, no copying—just fresh, well-researched work with proper citations, crafted by real experts. You can grab a plagiarism report to see it’s 95%+ original, giving you total peace of mind it’s one-of-a-kind and ready to impress.

175

200

500

1 in 4

Can You Use Any Citation Format?

Yep—APA, Chicago, Harvard, MLA, Turabian, you name it! Our writers customize every detail to fit your assignment’s needs, ensuring it meets academic standards down to the last footnote or bibliography entry. They’re pros at making your paper look sharp and compliant, no matter the style guide.

50

150

200

750

Can I Change My Order Details?

For sure—you’re not locked in! Chat with your writer anytime through our handy system to update instructions, tweak the focus, or toss in new specifics, and they’ll adjust on the fly, even if they’re mid-draft. It’s all about keeping your paper exactly how you want it, hassle-free.

1 in 3

75

125

200

How Do I Order a Paper?

It’s a breeze—submit your order online with a few clicks, then track progress with drafts as your writer brings it to life. Once it’s ready, download it from your account, review it, and release payment only when you’re totally satisfied—easy, affordable help whenever you need it. Plus, you can reach out to support 24/7 if you’ve got questions along the way!

1000

1 in 2

100

100

How Quick Can You Write?

Need it fast? We can whip up a top-quality paper in 24 hours—fully researched and polished, no corners cut. Just pick your deadline when you order, and we’ll hustle to make it happen, even for those nail-biting, last-minute turnarounds you didn’t see coming.

200

Sample

Dilution Factor

Volume (µl)

How Do You Match Professor Expectations?

We follow your rubric to a T—structure, evidence, tone. Editors refine it, ensuring it’s polished and ready to impress your prof.

Sample

Volume (µl)

Added Water

Final Volume (µl)

How Do You Edit My Work?

Send us your draft and goals—our editors enhance clarity, fix errors, and keep your style. You’ll get a pro-level paper fast.

Biopharmaceutical

1 in 5

20

80

100

Can You Brainstorm Topics?

Yep! We’ll suggest ideas tailored to your field—engaging and manageable. Pick one, and we’ll build it into a killer paper.

Concentration mg/ml

Absorbance 1

Absorbance 2

Absorbance 3

Average

0

Do You Offer Fast Edits?

Yes! Need a quick fix? Our editors can polish your paper in hours—perfect for tight deadlines and top grades.

0

0

0

0

25

0.085141386

Can You Start With an Outline?

Sure! We’ll sketch an outline for your approval first, ensuring the paper’s direction is spot-on before we write.

0.050185522

0.149322437

0.094883

125

0.014410967

0.117177903

0.100239697

0.077276

Can You Add Charts or Stats?

Definitely! Our writers can include data analysis or visuals—charts, graphs—making your paper sharp and evidence-rich.

250

0.051610414

0.087607308

0.174962059

0.104727

500

0.53853473

0.338323087

0.351367406

0.409408

What About Multi-Part Projects?

We’ve got it—each section delivered on time, cohesive and high-quality. We’ll manage the whole journey for you.

750

0.622078392

0.563260249

0.452668921

0.546003

Do You Adapt to International Rules?

Yes! UK, US, or Aussie standards—we’ll tailor your paper to fit your school’s norms perfectly.

1000

0.773383051

0.85954018

0.870560367

0.834495

What does a complex assignment mean?

If your assignment needs a writer with some niche know-how, we call it complex. For these, we tap into our pool of narrow-field specialists, who charge a bit more than our standard writers. That means we might add up to 20% to your original order price. Subjects like finance, architecture, engineering, IT, chemistry, physics, and a few others fall into this bucket—you’ll see a little note about it under the discipline field when you’re filling out the form. If you pick “Other” as your discipline, our support team will take a look too. If they think it’s tricky, that same 20% bump might apply. We’ll keep you in the loop either way!

Sample (x)

0.155851667

0.056575722

-0.105354519

Who is my writer? How can I communicate with him/her?

Our writers come from all corners of the globe, and we’re picky about who we bring on board. They’ve passed tough tests in English and their subject areas, and we’ve checked their IDs to confirm they’ve got a master’s or PhD. Plus, we run training sessions on formatting and academic writing to keep their skills sharp. You’ll get to chat with your writer through a handy messenger on your personal order page. We’ll shoot you an email when new messages pop up, but it’s a good idea to swing by your page now and then so you don’t miss anything important from them.

0.035691

Calculation:

Equation of the linear least square fit can be represented as outlined below.

Y = 0.0008 (X) – 0.0051

0.035691= 0.0008 (X) – 0.0051

X = 0.035691 + 0.0051/0.0008

X = 0.040791/0.0008

X = 50.98875

Taking the dilution factor into account we multiply by 5

The protein concentration is thus 50.98875 x 5 = 254.94mg/ml

Discussion:

The sample data had an anomaly, showing a negative reading on the third well. This is suggestive of contamination. The possibility of interference from the reagent can be ruled out because the standard was treated the same way and also taking into cognisance the fact that the experiment was not carried out under a non-denaturing condition. The likely cause could be due to dirt on the Microwell thus blocking out the necessary wavelength for the absorbance reading or the sampling pipette not delivering the right amount of reagent. The intensity of the colour change for the third well was observed to be less than the other two wells. There is also the possibility of the sample not being vortexed properly or sample settling to the bottom of tube.

As mentioned earlier there seem to be an anomaly with our absorbance reading and this can be validated from our standard curve as it is not quite linear and on this basis we cannot absolutely rely on the result of the experiment.

2.1 Size Exclusion Chromatography Experiment

The priming and purging of the UPLC instrument was carried out by the trainer as per the instruction manual. System suitability testing was then carried out to ensure that it is fit for purpose. The test serves to assure the reproducibility of the instrument and the method. It is a regulatory requirement which was mentioned in both the EU and US pharmacopoeias. The testing is important as it can allow for critical factors that could affect the performance of the instrument to be adjusted to meet the test criteria. Parameters such as the resolution, efficiency of the column, tailing factors, relative standard deviation etc. are used as criteria for comparison with regards to standards and test samples.

The table below details the results obtained from the system suitability testing, reference standard and our test sample. The UPLC system used in our experiment can be said to be fit for purpose taking into consideration, the system suitability test. The results obtained were within our test criteria. The resolution of the peak and standard deviation of the different retention time was less than 1 which as a rule of the thumb is quite acceptable.

Comparing the test sample to the reference standard one would not fail to notice that the first peak in the reference standard was a dimer while the second peak was a monomer but in our test sample the retention time of the first peak was really short showing evidence of a high molecular weight aggregation .Also from our result the second peak was our product dimer while the third peak was our product monomer. This result serves to highlight the mechanism of protein aggregation and the reason why it should be minimised as it impacts on the yield of the product and moreover it can affect the potency and therapeutic potential of the parenteral. It is also worth mentioning that sometimes early elution may not necessarily mean that there is aggregation, it could be for the simple reason that sometimes intrinsically unstructured proteins can elute so fast that they tend to behave like aggregates. A molecular weight comparison testing can be used to differentiate them.

From our experiment the test sample showed increase aggregation and this can be explained given the fact that the experiment was not carried out under a non-denaturing environment. The possibility of column contamination or buffer contamination can help to encourage aggregation. It is also important that samples should be free of extraneous particles during injection as this can also be a determining factor. The changing environment of the mobile phase can also be construed as a possible cause of the aggregation observed. The temperature of the instrument is another factor that can play a role and as we all know that the Arrhenius theory of a 10°C increase in temperature speeding up a reaction does not relate to proteins as it rather opens up the pathway of denaturation and aggregation.

Questions:

Estimation of protein concentration is important as we have to know the amount of protein in our final product after fermentation to know if the bioprocess has to be optimised with regards to the expected titre value. The concentration of the protein can also allow for the portioning of the product into the right dosage formulation, certain therapeutic proteins are required in a very high dosage form and their production can be sometimes targeted at a particular section of the population e.g. during an epidemic outbreak to ensure potency and biological activity. The knowledge of the concentration of proteins can also allow us to work out the economy of scale with regards to the profit margin taking into account, the expense incurred in research and development and other aspect of the production process. It is also important to estimate the amount of protein in our biomass so as to be able to optimise our subsequent purification steps. The estimation of the protein concentration can also give us an idea of product related impurities and those associated with the process.

Proteins are very complex molecules and are prone to several types of condition than cause instability from the starting stage of production to the end of their shelf life. Aggregation can be described in a layman’s term as the propensity for proteins to stick together under conditions such as a slight increase in temperature, pH, shear force, ionic strength of the solution they are contained in etc. Aggregation have been seen to cause delay in several novel biologics due to the debilitating effect on the health of the population that the drug is directed at and also in the context of compliance to regulatory authority as there is a specification to the amount of aggregates that can be allowed. It is extremely difficult if not impossible to totally eradicate aggregation from the process. It is worth mentioning however that the mechanism of aggregation is still subject to debate as it has not been fully understood. Aggregation can be reversible or irreversible depending on the stage it has attained as can be loss of primary structure.

The potency of biologics as we all know are normally related to them being in their native structure, in most instances aggregation leads to the loss of activity and moreover the overall yield of the biotherapeutic is greatly affected. Aggregation has also been known to spur immune response in patients that have been administered with protein therapeutics affected by aggregation this could be by way of the neutralisation of antibodies that helps to ensure the effectiveness of the drug. In a worst case scenario the immunogenic reaction can lead to incurable conditions such as seen in patients with pure red cell aplasia where the red blood cells are attacked and blood transfusion is needed for life. The route of administration of biologics is intravenous and the presence of aggregation especially those of very high molecular size can result in the blockage of blood vessels. It is thus very important that at each stage of our production testing should be carried out to check for aggregation.

Size exclusion chromatography is a purification system that exploits the molecular size of the compound of interest. Simply put it works just like a molecular sieve, smaller particles passes through the sieve which is the stationary phase and could be a bead coupled to a resin. The pore size of the beads are defined and on this basis it will only allow certain particle sizes to pass through while excluding those that are too large for the pore. The larger particles because they are not passing through the beads are thus excluded quickly, their retention time is thus said to be short. The smaller particles are retained longer while the larger particles earlier mentioned are eluted through the void volume. Different gels in use would typically have different pore sizes and can be used to determine the size of the molecules to be separated.

Despite all the numerous advantages of size exclusion chromatography which has made it the gold standard over the years for analysing protein aggregation there are still some limitation associated this method. The possibility of the stationary phase and the analyte reacting together can be sometimes rife thus leading to a longer retention time which serves to mimic the compound as being of low molecular size. The cost associated with running this type of separation technique can also be enormous due to the fact that large columns and eluents are required and this serves to add to the overall cost of the unit operation. In comparison to other modes of separation, size exclusion chromatography can be said to have an inherent low resolution as there is a limited range of molecular weight that can be separated as a result of dependence on the pore size of the beads in use. There is also the possibility of proteolytic degradation as the protein of interest can become targets for proteolytic enzymes still present in solution. The accuracy of this technique can sometimes come into question due to the fact some aggregates will remain in solution and as such would not be detected. Also taking into consideration the fact that larger molecular aggregate leaves the column through the void volume, there is also the possibility. The possibility of the polymer in use to degrade is also a drawback as this can occur at a very high flow rate. The high flow rate as mentioned earlier can degrade the polymer and it also has the ability of altering the geometry of the beads in use making the separation technique inefficient

Order|Paper Discounts