Sciencemadness Discussion Board

Lactoferrin COA confusion-help?

Lacto - 26-11-2018 at 03:15

Hi there,
Anybody willing to give me the benefit of their knowledge interpreting a COA?
I have received a quantity of lactoferrin. The COA states a purity of 98% and an iron saturation of 15%.
How would you interpret that?
Does it mean that 15% of the material is iron?
Or does it mean that of the two Fe pockets on the transferrin molecule, 15% are filled?
How would I calculate the iron content of one gram and it’s relation to the RDA?
Thanks!

Texium - 26-11-2018 at 08:14

It most certainly means that 15% of the iron binding sites are occupied. It would be impossible for it to be 15% iron by mass and also 98% pure!

To find the amount of iron per gram you would need to take the atomic mass of iron, divided by the molar mass of the protein, multiplied by the number of iron atoms that the protein can bind, multiplied by 0.15

(Hint: for an 80 kDa protein like this your answer will be very small)

Welcome to the forum

Lacto - 27-11-2018 at 17:03

Thank you! I really appreciate your help.
Ok,
so that would be:

55.845 u
/
80kDA
X
2
X
0.15

= 0.2094

Am I still in u at this point? Or am I some how already in grams? Thought I needed some avocado (sic) in here somewhere?!

Texium - 28-11-2018 at 07:00

No, you're not in any units- it's a mass/mass ratio. You've got the correct number, but somehow it's off by a few orders of magnitude... try again

Tsjerk - 28-11-2018 at 09:57

"55.845 u" can be read as "one mole of this stuff weighs 55.845 grams"; the factor 2 and 0.15 you probably understand.

Now for the 80 kDa; this is also a way to express how much a mole weighs, it is basically the same as the 55.845 used for iron, but the weight of a mole of iron and the weight of a mole of protein is multiple orders of a magnitude different. Try to find out what kDa stands for. One mole of protein is a lot heavier than a mole of iron.

Lacto - 30-11-2018 at 01:29

Ok I see it.
u = da, and we are using Kda,
So the magnitude error is three decimal places out giving instead:
0.0002094
And that is in grams so:
0.2094 milligrams or
209.4 micrograms
Per gram of lactoferrin.

RDA recommendations vary widely.
One source advises that men need to absorb 1mg daily from a dietary intake of between 8-20mg. This appears to be considering traditional elemental supplements, however at face value, iron for iron assuming 100% absorption, would require 5 grams of lactoferrin daily.

Does anyone have a handle on the mechanisms lactoferrin uses to regulate iron balance and from there a guess at an equivalent RDA for this form of iron supplementation?

Thanks.

Texium - 30-11-2018 at 10:07

What are you trying to do with this?

I'm really curious as to what someone who (forgive my assumption) appears to be a chemistry novice, would be wanting to do with a protein on a multiple kilogram scale...

Lacto - 5-12-2018 at 18:21

Hmmm.... unfortunately my interest was sparked by the same thing that drives most into a crash course in biochemistry, a family members cancer diagnosis.
Before anyone dives in, may I remind you: Pubmed is your friend.
From that initial oncological interest I have followed it to it’s use as a antibiotic replacement, viral defence, gut health, anti fungal, oral health, and am now interested in it’s superior effect as an anemia treatment.
It is actually a fascinating molecule, more so since the recent tech leap that now enables its more economical extraction in large quantities.
Many of the various research studies compare effectiveness of the appo (0%)and holo (100%) versions at the ends of the spectrum, but it’s maturally occurring ratio is 15-30% saturation, which is the sample I have.

Having figured out that the amount of Iron each gram contributes, I can now advise my family member that they can probably use it in place of Appo, without too much worry as to increasing their iron load. (Tumours like iron.) interestingly tho, it seems that two week prior supplementation with holo increases tumour reduction with chemo.

Down the rabbit hole.....

Metacelsus - 5-12-2018 at 23:57

Specifically, what kind of cancer is it? "Cancer" isn't just one disease, and treatments for one kind of cancer may not be effective against others. Specifically, lactoferrin may make breast cancer worse (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5470622/ ):
Quote:
Recently, it was also postulated that elevated levels of lactoferrin are associated with reduced expression of ERα and PR, and perhaps HER-2, and therefore could contribute to the development of triple negative breast cancer (TNBC) phenotypes [15]. It was found that lactoferrin efficiently downregulates ERα, PR, and HER-2, and the increased invasiveness and aggression of these breast cancer cell lines was attributed to the lactoferrin-endothelin axis.

Based on the several experiments by Ha et al. [15], it was found that persistent exposure of breast cancer cells to lactoferrin resulted in downregulation of the hormone receptors leading to the development of TNBC phenotypes.


(Also, I saw a recent paper you might be interested in about anticancer activity of mannose. It's blogged about here: http://blogs.sciencemag.org/pipeline/archives/2018/12/05/man... )

Lacto - 11-12-2018 at 18:52

Hi,
Thanks for your time on this.
The mannose link was particularly useful and appreciated. My family member has already transitioned to a ketogenic diet, so I will have to look into the mechanism by which mannose inhibits glucose availability to see if it will be a helpful addition in his case.

After reading the paper your quoted paper linked to, I am not convinced that BLf is a problem for breast cancer. What it seemed to say to me was that, in vitro, as in a pietri dish, Lf (human Lf) down regulates three of the receptor s that are currently targeted for therapeutic treatment for triple negative breast cancer, which I take to mean Lf makes current targeted pathway treatments less effective. Secondly Lf binds to ET1 which is another unhelpful mechanism of action, however it then states that bovine Lf does not bind to ET1, so no problem there as supplemental Lf is blf not human Lf. The paper you quote then goes on to say that there are contrary studies that give a positive effect for the use of blf in breast cancer.

Independent of all this, our particular issue is lung cancer, for which there are some interesting and compelling, murine (mouse based) effects for bLf, and I think a couple of in vivo (human) research studies also.

It’s a pleasure engaging with you.
Thank you.